CN105228627A

CN105228627A - Leucine and nicotinic acid reduce lipid level

Info

Publication number: CN105228627A
Application number: CN201480027603.9A
Authority: CN
Inventors: 麦克·泽梅尔; 安提亚·布鲁克鲍尔
Original assignee: Nusirt Sciences Inc
Current assignee: Nusirt Sciences Inc
Priority date: 2013-03-15
Filing date: 2014-03-13
Publication date: 2016-01-06
Anticipated expiration: 2034-03-13
Also published as: US9895357B2; ZA201606052B; US20170340615A1; PH12015502033A1; EP2968333A4; ZA201705790B; US20180214421A1; AU2014236687A1; JP6550370B2; IL240762A0; EP2968333B1; US9707213B2; WO2014152016A1; ZA201805629B; RU2015143836A; CA2902879A1; ZA201506481B; CN105228627B; JP2019065049A; US9408834B2

Abstract

There is provided herein the compositions, method and the test kit that can be used for treating the hyperlipemia patient's condition.Such compositions can nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite containing collaborative amount combine with leucine and/or leucine metabolite, containing or do not contain resveratrol.

Description

Leucine and nicotinic acid reduce lipid level

cross reference

This application claims the priority of the U.S. Provisional Patent Application numbers 61/800,363 submitted on March 15th, 2013, this provisional application is incorporated to herein with its entirety by reference.

Background technology

Dysbolismus is as hyperlipemia and obesity and present the remarkable burden to publilc health to relative influence that is healthy and mortality rate.Such as, body-mass index is defined as clinically more than 30kg/m ²obesity have impact on according to estimates 35.7% US adult population.In the U.S., obesity causes annual about 110,000-365,000 example dead according to estimates.Obesity can cause hyperlipemia, it is characterized in that blood flow comprises cholesterol, cholesteryl ester, phospholipid and triglyceride excessive at interior lipid, and diabetes, angiopathy, cancer, nephropathy, infectious disease, external cause, deliberate oneself's injury, neurological conditions and chronic lung disease (NEnglJMed2011 can be caused further; 364:829-841).According to estimates, the metabolism syndrome that experimenter shows central obesity and at least two kinds of other dysbolismus (as hypercholesterolemia, hypertension or diabetes) have impact on the U.S. population of 25%.

Hyperlipemia---a kind of symptom of obesity and other patient's condition, can treat with the multi-medicament comprising nicotinic acid.Nicotinic acid is a kind of form of vitamin B3 (nicotinic acid).When taking with high dose (1-3g/ days), nicotinic acid can treat hyperlipemia, because it can reduce TL, LDL, cholesterol, triglyceride and lipoprotein, or raises the HDL lipoprotein in blood flow.It can also reduce the progress of atheromatous plaque and the M & M of coronary heart disease.

But nicotinic acid may have significant side effect, therefore may normally bad tolerance.A kind of significant side effect may be serious cutaneous vasodilation and flare reaction, although therefore demonstrate safety and effectiveness well, seldom output (CarlsonLA.Nicotinicacid:thebroad-spectrumlipiddrug.A50th anniversaryreview.JIntMed2005 as prescription; 258:94-114).Although side effect slightly weakens in lasting (SR) and prolongation (ER) delivery formulations, side effect still lasts till the use being enough to limit drug.Therefore, there is the urgent needs for the side effect reducing nicotinic acid when not reducing its therapeutic effect.

Summary of the invention

This application provides the compositions of the hyperlipemia being used for the treatment of experimenter, method and test kit.Said composition, method and test kit can comprise the leucine and/or leucine metabolite that combine with nicotinic acid.The invention solves with negative side effect during high dose nicotinic acid treatment experimenter.

Compositions of the present invention can oral or by other approach as intravenous administration is used.Pill, tablet, capsule etc. can be comprised for oral compositions.

In one aspect, the invention provides and comprise at least about 250mg leucine and/or one or more leucine metabolite of about 25mg and the compositions at least about 1mg nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite.

In another aspect of this invention, provide the compositions comprised at least about 250mg leucine and/or one or more leucine metabolite of about 25mg and a certain amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, wherein said composition is substantially free of each in alanine, glycine, glutamic acid and proline.

In another, the present invention goes back providing package containing the compositions at least about 250mg leucine and/or one or more leucine metabolite of about 25mg and a certain amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, and wherein the described amount of nicotinic acid and/or nicotinic acid metabolite is not enough to when there is not component (a) reduce lipid content.

In another aspect of the invention, provide the compositions comprising a certain amount of leucine and/or one or more leucine metabolite and a certain amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, wherein use said composition to experimenter in need, further wherein compared to the dosage of only nicotinic acid with said composition in the lipid content reducing experimenter with identical effectiveness, the cutaneous vasodilation that said composition causes degree to reduce in used experimenter.

On the other hand, the present invention further provides and comprise leucine and/or one or more leucine metabolite and the compositions at least about 1mg nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, in wherein said compositions, component (a) is greater than about 20 with the mol ratio of (b).

In some embodiments, said composition as herein described comprises at least about 500mg leucine and/or at least about one or more leucine metabolite described in 200mg.In some embodiments, said composition comprises at least about 250mg leucine and/or one or more leucine metabolite of about 25mg.In some embodiments, the amount of leucine and/or one or more leucine metabolite is less than about 1g.In some embodiments, the amount of leucine and/or one or more leucine metabolite is less than 3g.

Compositions disclosed herein can be substantially free of nicotiamide.In some embodiments, said composition can be substantially free of nicotinic acid metabolite.In another embodiment, said composition can be substantially free of each in nicotinoyl coenzyme A, nitocinoylglycine (nicotinuricacid), NAMN, nicotinate adenine dinucleotide and nicotinamide adenine dinucleotide.In some embodiments, this nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite are nicotinic acid.

Compositions disclosed herein can be substantially free of leucine metabolite.In some embodiments, this leucine and/or one or more leucine metabolite are leucines.

In some cases, compositions as herein described can comprise a certain amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, and this amount may be less than about 1g.In some cases, the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can be less than about 250mg.In some embodiments, the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can be about 1-100mg.In some embodiments again, the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can be at least about 1mg.

In some embodiments, compositions disclosed herein can comprise a certain amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, and this amount can realize being less than the nicotinic acid of about 100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.In another aspect of this invention, the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize the nicotinic acid of about 10nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.In another, the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize the nicotinic acid of about 1-100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

In some cases, compositions disclosed herein can make the triglyceride levels of experimenter, T-CHOL or LDL level reduce at least about 5% effectively.In some cases, in said composition, the amount of component (a) and (b) can reduce the lipid content of described experimenter synergistically when being applied to experimenter.In some embodiments, the increase that the reduction that the body weight that the component (a) in said composition and component (b) can strengthen experimenter synergistically increases, the increase of the fat oxidation of experimenter or the Sirt1 of experimenter activate.

In some embodiments, when there is not leucine and/or one or more leucine metabolite, in compositions as herein described, the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite may be not enough to reduce lipid content.

In some cases, compositions disclosed herein can be included in food.

In some embodiments, a part for the leucine that comprises of compositions of the present invention and/or one or more leucine metabolite can be free form.In some embodiments, a part for this leucine and/or one or more leucine metabolite can be salt form.

More of the present invention in, said composition can comprise resveratrol further.

In some cases, said composition can be prepared for oral administration.In some cases, said composition can be tablet, capsule, pill, granule, Emulsion, gel, multiple beadlet, powder, suspension, liquid, semiliquid, semisolid, syrup, serosity or the chewable form be encapsulated in capsule.

In some cases, the component (a) comprised in compositions as herein described and component (b) can separately be packed.In some embodiments, component (a) and component (b) can mix.

Compositions as herein described can be substantially free of the every seed amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine.In some cases, said composition can be substantially free of the often kind of free amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine.In some cases, said composition can containing the often kind of free amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine being less than about 0.1%.In some embodiments, said composition can containing the non-leucine aminoacid being less than about 10%.

On the other hand, one or more leucine metabolite that can be included in compositions as herein described can be selected from ketone-isocaproic acid (KIC), alpha-hydroxy-isocaproic acid and HMB.In some embodiments, compositions disclosed herein can not comprise nicotiamide.

In another, compositions disclosed herein can comprise the therapeutic agent that one or more can reduce lipid accumulation further.In some embodiments, one or more therapeutic agents that can be included in compositions as herein described can be selected from HMG-CoA inhibitor, fibrate (fibrate), bile acid chelating agent, ezetimibe (ezetimibe), Lome he send (lomitapide), phytosterol (phytosterols), CETP antagonist, orlistat (orlistat) and combination in any thereof.

In some embodiments, in compositions as herein described, component (a) can be greater than about 20 with the mol ratio of (b).

In some embodiments, compositions as herein described can be formulated as unit dosage forms.

On the other hand, present invention also offers test kit, it comprises supplies in many days of the unit dose of compositions as herein described, and instructs the description using supplies in described many days within a period of time of many days.

In another, invention further provides the method for the total cholesterol level reduced in experimenter in need, it comprises uses compositions disclosed herein to realize the total cholesterol level in this experimenter to described experimenter.In some embodiments, present invention also offers the method for the total lipid content reduced in experimenter in need, it comprises uses compositions disclosed herein to realize the total lipid content in this experimenter to described experimenter.

In one aspect, present invention also offers the method for the total cholesterol level reduced in experimenter in need, it comprises the compositions comprising leucine and/or one or more leucine metabolite and a certain amount of nicotinic acid and/or nicotinic acid metabolite using doses to described experimenter, to realize the total cholesterol level in this experimenter.

In some embodiments, the nicotinic acid that can use in method disclosed herein and/or the amount of nicotinamide riboside and/or nicotinic acid metabolite can be less than about 250mg.In some embodiments, the nicotinic acid that can use in method disclosed herein and/or the amount of nicotinamide riboside and/or nicotinic acid metabolite can be less than about 100mg.In some embodiments, the nicotinic acid that can use in method described herein and/or the amount of nicotinamide riboside and/or nicotinic acid metabolite can be less than about 25mg.In some embodiments, the nicotinic acid that can use in method described herein and/or the amount of nicotinamide riboside and/or nicotinic acid metabolite can be less than about 10mg.In some embodiments, the dosage of the compositions that can use in method disclosed herein can be unit dose.

In another aspect of this invention, disclose the method for the side effect reducing nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, wherein the feature of this side effect can be the increase of the cutaneous vasodilation used in the experimenter of nicotinic acid and/or nicotinamide riboside and/or these one or more nicotinic acid metabolite, and the method comprises the compositions using leucine and/or one or more leucine metabolite including effective amount to experimenter, it can be used with nicotinic acid and/or one or more nicotinic acid metabolite.

In some embodiments, method as herein described can comprise Orally administered said composition.

In some cases, the effective dose of the leucine used in method as herein described and/or leucine metabolite can comprise at least about 500mg leucine and/or at least about one or more leucine metabolite described in 200mg.In some cases, the effective dose of leucine and/or leucine metabolite can comprise at least about 250mg leucine and/or at least about one or more leucine metabolite described in 25mg.

In some embodiments, method disclosed herein can relate to nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, if used separately, it is sub-therapeutic dose.In some embodiments, the nicotinic acid used in method as herein described and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can be the amounts being less than about 1g.In some embodiments, the nicotinic acid used in method disclosed herein and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can be the amounts being less than about 250mg.In some embodiments, the amount of the nicotinic acid used in method as herein described and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can be about 1-100mg.

In some cases, method as herein described can use a certain amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, and this amount can realize the nicotinic acid of about 1-100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

In still another aspect of the invention, the method for the atheromatous plaque size reduced in experimenter in need is provided.The method comprises the compositions comprising leucine and/or one or more leucine metabolite and a certain amount of nicotinic acid and/or nicotinic acid metabolite using doses to described experimenter, to realize the total atheromatous plaque size in this experimenter.

In some embodiments, the nicotinic acid that can use in method as herein described and/or the amount of nicotinamide riboside and/or nicotinic acid metabolite can be less than about 250mg.In some embodiments, the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can be about 1-100mg.In some embodiments, the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can be less than about 25mg.

In some cases, method as herein described can comprise to described experimenter's applying said compositions at least about 1 year.

In some embodiments, the dosage of the compositions used in method as herein described can be unit dose.

quote and be incorporated to

The all publications mentioned in this description, patent and patent application are all incorporated to by reference at this, and its degree is as pointed out that each independent publication, patent or patent application are incorporated to by reference particularly and individually.

Accompanying drawing explanation

Novel feature of the present invention is specifically set forth in the appended claims.By reference to the following the detailed description and the accompanying drawings set forth the illustrative embodiment using the principle of the invention wherein, the better understanding to the features and advantages of the present invention will be obtained, in the accompanying drawings:

Fig. 1 shows the chemical constitution of nicotinic acid and nicotinamide riboside.

Fig. 2 shows the impact that nicotinic acid and leucine and/or resveratrol activate the Sirt1 in C2C12 myotube.NA refers to nicotinic acid; Leu refers to leucine; R refers to resveratrol.*p<0.05；**p＝0.0001。The % that data are expressed as apart from control value changes.

Fig. 3 shows the impact on the P-AMPK/AMPK ratio in 3T3-L1 adipose cell of nicotinic acid and leucine and/or resveratrol.NA refers to nicotinic acid; Leu refers to leucine; R refers to resveratrol.*p<0.01。The % that data are expressed as apart from control value changes.

Fig. 4 shows the impact (* p=0.012) of leucine (0.5mM)/nicotinic acid (10nM) on the lipid level in Caenorhabditis elegans (C.elegans).NA refers to nicotinic acid; Leu refers to leucine.

Fig. 5 shows in ldl receptor knock-out mice, with being added into western diet (WesternDiet, WD) leucine (Leu in, 24g/kg diet), Leu (24g/kg diet)+nicotinic acid (NA, 50mg/kg diet), Leu (24g/kg diet)+NA (250mg/kg diet) and NA (1,000mg/kg diet) processes the impact of surrounding on total plasma cholesterol.

Fig. 6 shows in ldl receptor knock-out mice, with the leucine (Leu be added in western diet (WD), 24g/kg diet), Leu (24g/kg diet)+nicotinic acid (NA, 50mg/kg diet), Leu (24g/kg diet)+NA (250mg/kg diet) and NA (1,000mg/kg diet) processes the impact of surrounding on total plasma cholesterol ester.

Fig. 7 shows in ldl receptor knock-out mice, with the leucine (Leu be added in western diet (WD), 24g/kg diet), Leu (24g/kg diet)+nicotinic acid (NA, 50mg/kg diet), Leu (24g/kg diet)+NA (250mg/kg diet) and NA (1,000mg/kg diet) processes the impact of surrounding on plasma triglyceride.

Fig. 8 shows nicotinic acid and leucine to the impact in the life-span of Caenorhabditis elegans.NA refers to nicotinic acid; Leu refers to leucine.*p<0.0001。Data are expressed as the % survival of passing in time.

Detailed description of the invention

Term used herein is only used to the object describing specific embodiments, and also not intended to be limiting the present invention.As used herein, singulative " ", " one " and " being somebody's turn to do " are also intended to comprise plural form, unless the context clearly indicates otherwise.In addition, " comprise " with regard to term, " comprising ", " having ", " having ", " with " or the use of its version in detailed description of the invention and/or claim, these terms are intended to be similar to the mode that term " contains " and are included.

Term " about " or " approximately " are meant to as one of ordinary skill in the identified, and for being specifically worth in acceptable range of error, this partly will depend on how this value is measured or determine, i.e. the restriction of measuring system.Such as, according to the practice of this area, " about " may mean 1 or more than 1 standard deviation within.Or " about " may mean reaching 20%, can reach 10%, can reaching the scope that 5% maybe can reach 1% of set-point.Or particularly for biosystem or process, this term may mean in the order of magnitude of value, preferably within 5 times, more preferably within 2 times.When describing particular value in the application and claims, unless otherwise stated, being meant in the acceptable range of error of particular value of term " about " should be supposed.

As used herein, term " experimenter " or " individuality " comprise mammal.Mammiferous limiting examples comprises people and mice, comprises transgenic and nontransgenic mice.Method described herein can be used for human treatment, clinical front and veterinary's application.In some embodiments, experimenter is mammal, and in some embodiments, experimenter behaves.Other mammals include but not limited to, ape, chimpanzee, orangutan, monkey; Performing animal (house pet), as Canis familiaris L., cat, Cavia porcellus, hamster, mice, rat, rabbit and ferret; Domesticated animal, as cattle, Babalus bubalis L., wild ox, horse, donkey, pig, sheep and goat; Or usually at the zoo in the wild animal seen, such as Bears, lion, tiger, leopard, resemble, river horse, rhinoceros, giraffe, Saigae Tataricae, sloth, gazelle, zebra, gnu, prairie dog, koala, kangaroo, panda, giant panda, hyena, sea dog, sea lion and walrus.

Term administering " and " administration " be defined as providing compositions by approach known in the art to experimenter, this approach include but not limited to intravenous, intra-arterial, oral, parenteral, buccal, locally, percutaneous, rectum, intramuscular, in subcutaneous, bone, through mucous membrane or Intraperitoneal medication approach.In some embodiment of the application, the oral route using compositions may be preferred.

As used herein, " medicament " or " bioactivator " refers to biology, pharmacy or chemical compound or other parts.Nonrestrictive example comprises simple or complicated organic or inorganic molecule, peptide, protein, peptide nucleic acid(PNA) (PNA), oligonucleotide (comprise, such as, fit and polynucleotide), antibody, antibody derivatives, antibody fragment, vitamin derivative, sugar, toxin or chemotherapy compound.Various compound can be synthesized, such as, micromolecule and oligomer (such as, oligopeptide and oligonucleotide), and based on the anthropogenics of multiple core texture.In addition, multiple natural origin also can provide compound for screening, such as plant or animal extracts etc.Those of skill in the art will readily recognize that, the structural property for medicament of the present invention does not limit.

Term " effective dose " or " treatment effective dose " refer to the amount of compound as herein described, and it is enough to the application realizing expection, include but not limited to as undefined disease or patient's condition treatment.Treatment effective dose can according to the application (external or body in) of expection or the experimenter treated and disease condition (such as, the order of severity, administering mode etc. of the body weight of experimenter and age, the state of an illness) and different, and it easily can be determined by those of ordinary skill in the art.This term is also applicable to will the dosage of particular responses (such as, the minimizing of propagation or the downward of target protein activity) in inducing target cell.Concrete dosage by according to selected particular compound, the dosage regimen that follow, whether it co-administered with other compounds, delivery time, its tissue be administered to and carry its physical delivery system and change.

Term as used herein " energy metabolism " refers to the conversion of the energy of biochemical reaction in comitative aspect, and this biochemical reaction comprises cellular metabolism and mitochondrion biology occurs.Energy metabolism can quantize with many measuring methods described herein, such as but not limited to, to lose weight, fat loss, insulin sensitivity, fatty acid oxidation, glucose utilization, content of triglyceride, Sirt1 expression, AMPK expression, oxidative stress and mitochondrion Biomass.

When being applied to component of the present invention, such as PDE5 inhibitor, include but not limited to nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, when leucine and leucine metabolite (as HMB) and resveratrol, term " separation " refers to the goods of the material lacking other components of at least some, and described component also may be present in this material or the natural existence of similar substance or obtain part at first.Therefore, such as, the material of separation is prepared by using purification technique this material of enrichment from the mixture of source.Enrichment can be measured based on absolute magnitude, the weight of the solution of such as per unit volume, or its can relative to exist in the mixture of source second, the material of potential interference measures.The more and more higher enrichment of embodiment of the present invention is preferred gradually.Therefore, such as, 2 times of enrichments are preferred, and 10 times of enrichments are preferred, and 100 times of enrichments are more preferably, and 1000 times of enrichments are even more preferably.Material can also pass through man-made assembly process (as passed through chemosynthesis) to be provided with the state be separated.

" the sub-therapeutic dose " of medicament, activator or therapeutic agent is such amount: it is less than this medicament, activator or therapeutic agent for expecting the effective dose applied, but when another kind of medicament or therapeutic agent with effective dose or sub-therapeutic dose, due to the side effect of the cooperative effect in the useful effect of such as gained and/or minimizing, the result of expectation can be produced.

" collaborative " or " working in coordination with " effect can be such, and it makes one or more effects of coupling compositions be greater than one or more effects of often kind of independent component, or it can be greater than one or more effect sums of often kind of independent component.Cooperative effect can than be used alone wherein a kind of component time approximately high or be greater than about 10%, 20%, 30%, 50%, 75%, 100%, 110%, 120%, 150%, 200%, 250%, 350% or 500% or even more to the effect of experimenter or the synergistic effect when often kind of component is used respectively.This effect can be any measurable effect described here.

Term as used herein " is substantially free of " compositions referring to and have and be less than about 10%, be less than about 5%, be less than about 1%, be less than about 0.5%, be less than 0.1% or less specific components.Such as, being substantially free of the amino acid whose compositions of non-branched can containing the non-branched amino acid lysine being less than about 1%.Percentage ratio can be defined as the percentage ratio of the percentage ratio of total composition or the subset of compositions.Such as, be substantially free of the amino acid whose compositions of non-branched and can have the non-branched aminoacid being less than 1%, its percentage ratio as total composition or as the amino acid whose percentage ratio in said composition.Percentage ratio can be quality, mole or percent by volume.

Term " clinical significance " or " significant clinically " expression are considered to the behavior and the symptom that exceed normal range, and are characterized in puzzlement and the infringement of daily life function.Such as, significant cutaneous vasodilation is enough to cause patient about the level of main suit of discomfort (comprising flushing, pruritus and/or tingling) being secondary to acute vasodilatation by being clinically.The level of cutaneous vasodilation also can be measured by any method known in the art, as people such as SaumetJ.L., " Non-invasivemeasurementofskinbloodflow:comparisonbetween plethysmography, laser-Dopplerflowmeterandheatthermalclearancemethod " Int.J.Microcirc.Clin.Exp.1986; The method of Laser-Doppler effusion meter is comprised disclosed in 5:73 – 83.The significant clinically level of cutaneous vasodilation also can be statistically evident level.The significant clinically level of cutaneous vasodilation also can be statistically insignificant level.

Term " lipid content " or " lipid level " refer in the lipid of experimenter's in-vivo measurement or the content of lipoprotein molecule or level.It can be the concentration of the lipid molecular in circulation blood flow, or the total amount of body fat.Lipid or lipoprotein molecule can comprise triglyceride, cholesterol, LDL or HDL.

compositions

Compositions of the present invention comprises the combination of (i) leucine and/or one or more leucine metabolite and (ii) nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite.Said composition can be used for treating hyperlipemia.Said composition can comprise resveratrol further or one or more can reduce the therapeutic agent of lipid level.The combination of these components may be used for reducing lipid content, reduces total cholesterol level, reduces LDL level, reduces triglyceride levels or improve HDL level.In some embodiments, prepare these components to provide cooperative effect, include but not limited to the further reduction of fat content or the reduction of dosage, thus cause reducing the side effect of experimenter.This combination can reduce lipid content especially effectively, simultaneously compared at the dosage reducing the only nicotinic acid with said composition in lipid content with identical effectiveness, causes the cutaneous vasodilation that degree reduces in experimenter.When there is not leucine and/or one or more leucine metabolite, the amount of the nicotinic acid in said composition and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can be sub-therapeutic dose.

In one embodiment, compositions of the present invention comprises leucine and/or one or more leucine metabolite; And nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, wherein said composition comprises at least about 250mg leucine and/or at least about one or more leucine metabolite described in 25mg, and further wherein said composition comprise at least about 1mg nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite.

In another embodiment, compositions of the present invention comprises leucine and/or one or more leucine metabolite; And nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, wherein said composition comprise at least about 250mg leucine and/or at least about 10,20,25,30,35,40,45,50,55, one or more leucine metabolite described in 60mg, and further wherein said composition be substantially free of each in the aminoacid including but not limited to alanine, glycine, glutamic acid and proline.

In yet another embodiment, compositions of the present invention comprises leucine and/or one or more leucine metabolite; And a certain amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, wherein said composition comprises at least about 250mg leucine and/or at least about one or more leucine metabolite described in 25mg, and further wherein when there is not leucine and/or one or more leucine metabolite, the quantity not sufficient of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite to show therapeutic effect, as reduce lipid content.In some embodiments, when not using together with leucine and/or one or more leucine metabolite, the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is sub-therapeutic doses.

In still another embodiment, compositions of the present invention comprises leucine and/or one or more leucine metabolite; And nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, wherein said composition reduces the lipid content in experimenter in need effectively, simultaneously compared at the dosage reducing the only nicotinic acid with said composition in lipid content with identical effectiveness, the cutaneous vasodilation causing degree to reduce in this experimenter.In some embodiments, said composition reduces the lipid content in experimenter in need effectively, and can not cause significant cutaneous vasodilation clinically.

In another embodiment, compositions of the present invention comprises (a) leucine and/or one or more leucine metabolite; (b) nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, wherein the mass ratio of (a) and (b) is at least about 10,15,20,25,30,35,40,45,50,55,60,65,70,75 or 100, and wherein said composition comprises at least about this nicotinic acid of 1mg and/or nicotinamide riboside and/or nicotinic acid metabolite.As described herein, can provide sub-therapeutic administratp at least about the nicotinic acid of 1mg and/or the dosage of nicotinamide riboside and/or nicotinic acid metabolite, it can be effective when combining with the leucine or leucine metabolite of enough mass ratioes.

In some embodiments, compositions of the present invention comprises (a) leucine and/or one or more leucine metabolite; And (b) nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, wherein component (a) and component (b) have cooperative effect.This cooperative effect can be the body weight strengthening experimenter synergistically increase reduction, the reduction of lipid content, the reduction of LDL level, the raising of HDL level, the reduction of cholesterol levels, the reduction of triglyceride levels, the fat oxidation of experimenter increase, or the increase that the Sirt1 in experimenter activates.

In one aspect of the invention, compositions of the present invention comprises (a) leucine and/or one or more leucines; And (b) nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite.Said composition can comprise further at least about 0.01,0.05,0.1,0.5 or 1 μ g resveratrol.

In yet another embodiment, compositions of the present invention comprises (a) leucine and/or one or more leucine metabolite; And (b) nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, wherein (b) exists with the amount effectively reaching the cyclical level of about 1-100nM nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite in experimenter.In some embodiments, the cyclical level of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is 10nM.These target circulation levels have proved corresponding to described herein the treatment concentration (see embodiment) the hyperlipemia patient's condition being provided to beneficial effect in experimenter.

nicotinic acid, nicotinamide riboside and nicotinic acid metabolite

The invention provides the compositions comprising nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite.This nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can use in a free form.Term " dissociates ", when mention in this article component in conjunction with time, represent that this component is not introduced comparatively in macromolecular complex.In some embodiments, this nicotinic acid can be included in nicotinic acid.This nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can be salt forms.

In some embodiments, said composition can be substantially free of nicotiamide and/or nicotiamide metabolite.This nicotiamide and/or nicotiamide metabolite can offset the effect of nicotinic acid or nicotinamide riboside.Nicotiamide may be harmful to liver at high doses (disclosed in http://www.livestrong.com/article/448906-therapeutic-levels-of-niacin-to-lower-cholesterol-levels/#ixzz2NO3KhDZu).The quality of nicotiamide and/or nicotiamide metabolite or mole can be less than about 0.01%, 0.1%, 0.5%, 1%, 2%, 5% or 10% of total composition.The quality of nicotiamide and/or nicotiamide metabolite or mole can be less than about 0.01%, 0.1%, 0.5%, 1%, 2%, 5% or 10% of total composition.

Not limiting by theoretical, the absorption of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can reduce lipid content, reduces triglyceride levels, reduces LDL level, reduces total cholesterol level or improve HDL level.The absorption of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can also improve fat oxidation or stimulate defying age enzyme (sirtuin) intracellular signaling, comprises the activation improving Sirt1 and Sirt3.In some embodiments, arbitrary composition described herein can comprise the salt of nicotinic acid, derivant, metabolite, catabolite, anobolite, precursor and analog.Such as, this metabolite can comprise nicotinoyl coenzyme A, nitocinoylglycine, NAMN, nicotinate adenine dinucleotide or nicotinamide adenine dinucleotide.In some embodiments, said composition can not comprise nicotiamide.In some embodiments, said composition comprises nicotiamide.In some embodiments, said composition can be substantially free of nicotinic acid metabolite.

leucine and leucine metabolite

The invention provides the compositions comprising leucine and/or leucine metabolite.This leucine and/or leucine metabolite can use in a free form.Term " dissociates ", when being combined with component in this article, represents that this component is not introduced comparatively in macromolecular complex.Such as, compositions can comprise and not introduce free leucine in protein or free HMB.This leucine can be L-Leu.This leucine and/or leucine metabolite can be salt forms.

Not limiting by theoretical, branched-chain amino acid can stimulate defying age enzyme intracellular signaling as leucic picked-up, comprises Sirt1 and Sirt3, and AMPK intracellular signaling, and wherein one or more can advantageously regulate inflammatory cytokine to compose.In some embodiments, arbitrary composition described herein can comprise leucic salt, derivant, metabolite, catabolite, anobolite, precursor and analog.Such as, this metabolite can comprise HMB (HMB), ketone-isocaproic acid (KIC) and ketoisocaproate salt.HMB can be various ways, comprises HMB hydrate of calcium.

In certain embodiments of the invention, containing (or eliminating), one or more are not selected from the aminoacid of lysine, glutamic acid, proline, arginine, valine, isoleucine, aspartic acid, agedoite, glycine, threonine, serine, phenylalanine, tyrosine, histidine, alanine, tryptophan, methionine, glutamine, taurine, carnitine, cystine and cysteine to make them can to prepare arbitrary composition disclosed herein.

In some embodiments, said composition can be substantially free of one or more or whole non-leucine aminoacid.Such as, said composition can not contain alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and/or valine.

Said composition can be substantially free of free non-leucine aminoacid, as alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and/or valine.

In some embodiments, said composition can be substantially free of one or more or all non-branched or non-leucine aminoacid.Such as, said composition can not contain alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan and/or tyrosine.In some embodiments, said composition can be substantially free of isoleucine and/or valine.Compositions of the present invention can be substantially free of single amino acids alanine, glycine, glutamic acid and proline.Compositions of the present invention can be substantially free of in single amino acids alanine, glycine, glutamic acid and proline one or more.Compositions of the present invention can be substantially free of alanine.Compositions of the present invention can be substantially free of glycine.Compositions of the present invention can be substantially free of valine.Said composition can be substantially free of any non-branched aminoacid.The amino acid whose quality of non-branched or mole can be less than total composition or in said composition total amino acids about 0.01%, 0.1%, 0.5%, 1%, 2%, 5% or 10%.The amino acid whose quality of non-leucine or mole can be less than total composition or in said composition total amino acids about 0.01%, 0.1%, 0.5%, 1%, 2%, 5% or 10%.

For the sake of clarity, aminoacid as herein described can be the whole amino acid existed with its free form or salt form.Such as, compositions of the present invention can be substantially free of free amino acid, as alanine, glycine, glutamic acid and proline.Non-branched aminoacid, any aminoacid or the amino acid whose quality of any non-leucine or mole can be less than total composition, in said composition total amino acids or in said composition total free amino acid about 0.01%, 0.1%, 0.5%, 1%, 2%, 5% or 10%.

therapeutic agent

Compositions of the present invention can comprise one or more forms of pharmacologically active agents except nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite or therapeutic agent further.This therapeutic agent or forms of pharmacologically active agents can be any medicaments known in the art.Such as, this coupling compositions can comprise the agent of pharmaceutical active hyperlipidemia further, or to lipid content also effective dietary supplement.This hyperlipidemia agent can be medicinal preparation for oral administration or injectable medicament.This hyperlipidemia agent can be the sub-therapeutic dose reducing total lipid content or triglyceride levels, LDL or cholesterol levels or raising HDL level.The type of hyperlipidemia agent known in the art can include but not limited to HMG-CoA inhibitor (or statins), fibrate, nicotinic acid, bile acid chelating agent (resin), cholesterol absorption inhibitor (ezetimibe), his group, phytosterol, orlistat etc. of Lome.The hyperlipidemia agent of statin type can include but not limited to: atorvastatin, fluvastatin, spectrum cut down statin, lovastatin, simvastatin, Pitavastatin, cerivastatin, Rosuvastatin or lovastatin/nicotinic acid ER.Cholesterol absorption inhibitor can include but not limited to the combination of ezetimibe and ezetimibe and simvastatin.The hyperlipidemia agent of the special type of shellfish can include but not limited to: gemfibrozil, fenofibrate, fenofibric acid, clofibrate or micronized fenofibrate.Bile acid chelating agent can include but not limited to: colestipol, colestyramine or colesevelam.The hyperlipidemia agent of other types can comprise dextrothyroxine sodium or eicosapentaenoic acid (icosapent).There is provided these examples only for purposes of discussion, and be intended to prove that range of applicability of the present invention is wide in numerous medicines.And do not mean that and limit the scope of the invention by any way.

Compositions of the present invention can comprise the therapeutic agent that one or more are medical herbs and/or supplement further.This medical herbs and/or supplement can have uncertified therapeutic effect in science.The example of this medical herbs and/or supplement includes but not limited to: Brazilian Acai (Acai), alfalfa, Aloe, aloe vera (AloeVera), Aristolochic Acid, Asia Radix Ginseng, the Radix Astragali (Astragalus), Bacillus coagulans (Bacilluscoagulans), Semen daturae, beta-carotene, bacillus bifidus (Bifidobacteria), Fructus Rubi, Pericarpium Citri tangerinae, biotin, bigarabe, Radix vernoniae asperae (BlackCohosh), Radix vernoniae asperae, black Herba Plantaginis, black tea, Fucus Vesiculosus, try to cover up flower (Blessedthistle), golden Semen Plantaginis (Blondpsyllium), blue berry, blue-green alge, boron, bromelain, coltsfoot (Butterbur), calcium, Flos Inulae, Cancell/Cantron/Protocel, cartilage (cattle and shark), Cortex Cinnamomi (Cassiacinnamon), Radix Ranunculi Ternati, Flos Chrysanthemi, the holy and pure certain kind of berries, chondroitin sulfate, chromium, Cortex cinnamomi japonici (Ramulus Cinnamomi), Flos Caryophylli, coenzyme q-10, collargol product, cranberry, creatine, Herba Taraxaci, Herba Taraxaci, Harpagophytum procumbens (Devil'sclaw), DHEA, Radix Angelicae Sinensis, Echinacea Species, Herba Ephedrae, Essiac/Flor-elite, Eucalyptus, Sambucusnigra (Ramulus Sambuci Williamsii), vicum album, Radix Oenotherae erythrosepalae oil, Semen Trigonellae, Staggerweed, fish oil, Semen Lini, Semen Lini oil, folate, folic acid, Bulbus Allii, Rhizoma Zingiberis Recens, Semen Ginkgo, Radix Ginseng, glucosamine hydrochloride, glucosamine sulfate, goldenseal (Goldenseal), Semen Vitis viniferae extract, green tea, Fructus Crataegi, Hoodia Cactus (Hoodia), Aesculus chinensis Bunge, Herba Equiseti Hiemalis (Horsetail), Hydrazinium sulfate, iodine, ferrum, slips (Kava), lactobacillus (Lactobacillus), amygdaloside/amygdalin, L-arginine, lavandula angustifolia, licorice (Licorice), Fructus Lycii, lycopene, magnesium, manganese, melatonin, Herba Silybi mariani, Herba Visci extract, Radix Morindae Officinalis (Noni), probiotic oral (OralProbiotics), pantothenic acid (vitamin B5), Herba Passiflorae Caeruleae, PC-SPES, Oleum menthae, Herba Menthae, phosphate, Punica granatum L., propolis (Propolis), pycnogenol, pyridoxol (vitamin B6), red Herba Trifolii Pratentis, Rhodothece glutinis, riboflavin (vitamin B2), ocean, Rome chrysanthemum, cloth Laplace yeast (Saccharomycesboulardii), SAMe (SAMe), Salvia japonica Thunb. (Sage), sabal, vegetable/Sun the soup selected, selenium, Folium Sennae (Senna), Semen sojae atricolor, Herba Hyperici perforati (St.John'sWort), Fructus Citri sinensis elite, tea tree oil, thiamine (vitamin B1), Radix Tripterygii Wilfordii (ThunderGodVine), Rhizoma Curcumae Longae (Turmeric), Rhizoma et radix valerianae (Valerian), vitamin A, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, dioscorea japonica, yohimbe (Yohimbe), zinc or 5-HTP.

The amount of the medicament used in coupling compositions described herein or arbitrarily other components can be use with sub-therapeutic dose.In some embodiments, the medicament of sub-therapeutic dose or component is used can to reduce the side effect of medicament.The use of sub-therapeutic dose still can be effective, particularly when with other medicaments or component is collaborative use time.

Medicament or the component of sub-therapeutic dose can be such, measure lower than being considered to curative amount to make it.Such as, the specific administration level of FDA guide possibility recommended therapy particular condition, and sub-therapeutic dose is exactly any level of the dose level lower than FDA suggestion.Sub-therapeutic dose can lower by about 1% than the amount being considered to therapeutic dose, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 50%, 75%, 90% or 95%.Can to individual subjects or in groups experimenter assess therapeutic dose.Experimenter can be whole potential experimenters or have the experimenter of special characteristic as age, body weight, race, sex or physiologically active level in groups.

When using separately nicotinic acid to reduce lipid content, doctor suggestion initial dose be 1000-3000mg every day, when using together with leucine and/or leucine metabolite, experimenter's specific dosage scope be every day 1mg to maximum 1000mg.Can be determined by carrying out titration to dosage and measuring therapeutic response by clinicist the specific administration amount of experimenter.Therapeutic administratp level can by measuring fasting plasma cholesterol when not causing significant cutaneous vasodilation clinically and LDL level is determined.Sub-therapeutic dose can be any level lower than the niacin dosage recommended.Such as, if the therapeutic administratp level of experimenter is confirmed as 700mg every day, then the dosage of 600mg is exactly sub-therapeutic dose.Or, sub-therapeutic dose can relative to one group of experimenter but not single experimenter determine.Such as, if for the experimenter of body weight more than 300lbs, the mean treatment amount of nicotinic acid, nicotinamide riboside or nicotinic acid metabolite is 2000mg, and so sub-therapeutic dose can be any amount lower than 2000mg.In some embodiments, this dosage can by include but not limited to the doctor of patient, nurse, nutritionist, pharmacists or other health care professional health care provider recommend.Health care professional can comprise the individual relevant to healthcare system or entity.The example of health care professional can comprise surgeon, dentist, audiologist, speech pathologists, internist's (comprising general practitioner and specialist), Physician's Assistant, nurse, midwives, pharmacist/pharmacists, nutritionist, therapist, psychologist, physiotherapist, phlebotomist, occupational therapist, optometrist, ridge is cured, clinical worker, emergency medical service technical staff, nursing staff, medical laboratory person, radiologic technologist, medical prostheses technician social worker, and numerous passing through is trained to provide other human resourcess of the Health Care Services of certain type.

For nicotinic acid, nicotinamide riboside or nicotinic acid metabolite, the treatment effect level of nicotinic acid, nicotinamide riboside, nicotinic acid metabolite can be the cyclical level of about 1-100nM.The sub-treatment level of nicotinic acid, nicotinamide riboside or nicotinic acid metabolite self or combination in any can be at least about, lower than about or higher than about 1,2.5,5,10,15,20,25,30,35,40,45,50,60,70,80,90 or any cyclical level of 100nM.Preparation is used for the sub-treatment level of nicotinic acid in the present composition of administration and/or nicotinamide riboside and/or nicotinic acid metabolite can lower than about 1,5,10,20,30,50,60,70,80,90,100,125,150,175,200,250,300,350,400,450,500,600,700,750,800,900 or 1000mg nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite.

Any component described herein, comprise leucine, HMB, KIC, nicotinic acid, nicotinamide riboside and resveratrol, can in the compositions of the present invention in a free form, unpack format, from natural origin purification and/or from synthesis source purification or preparation form use.Natural origin can be animal origin or plant origin.This component can be pure extremely at least about 95%, 97%, 99%, 99.5%, 99.9%, 99.99% or 99.999%.

dosage

The invention provides the compositions of the combination as the component be separated, the component of this separation is separated, as leucine, leucine metabolite, as HMB, nicotinic acid, nicotinamide riboside and/or resveratrol from one or more sources.The invention provides and be rich in leucine, leucine metabolite as the compositions of HMB, nicotinic acid and/or nicotinamide riboside and/or resveratrol.This component can be separated from natural origin or by the preparation of synthesis source, then enrichment is to improve the purity of this component.In addition, leucine can be separated from natural origin, then carries out enrichment by one or more separation methods.Be separated and the component of enrichment, as leucine, then can carry out combining and preparing for being applied to experimenter.

In some embodiments, compositions comprises a certain amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite.The amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can be sub-therapeutic dose, and/or with one or more other compounds in said composition or with said composition simultaneously or the collaborative amount of one or more compounds used at similar time.In some embodiments, nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite are used with low dosage, middle dosage or high dose, this describe the relation between two kinds of dosage, and usually do not limit any concrete dosage range.Said composition can be used, to make the said composition using selected total daily dose to this experimenter to experimenter.Total daily dose can be determined by the summation of the dosage used in 24 hours sections.

Can be that the dosage of unit dose can comprise approximately, more than about or be less than the leucine of about 200,250,400,500,550,600,700,800,900,1000,1100,1250,1300 or more mg.This leucine can be free leucine.In some embodiments, unit dose can comprise and to dissociate leucine at least about 1000mg.Said composition can comprise about 10-1250,200-1250 or 500-1250mg leucine.Can be that the dosage of unit dose can comprise approximately, more than about or be less than the leucine metabolite of about 10,15,20,25,30,35,40,45,50,100,200,250,400,500,550,600,700,800,900,1000,1250,1300 or more mg, as HMB or KIC.This leucine metabolite can be free leucine metabolite.Said composition can comprise about 10-900,50-750 or 400-650mg leucine metabolite, as HMB or KIC.In some embodiments, unit dose can comprise and to dissociate HMB at least about 400mg.The amount of leucine as herein described and leucine metabolite can every day or simultaneously use.Amount as herein described can use with a dosage or every day with multiple dosage separate administration.

In some embodiments, leucic daily dose can be approximately, be less than about or be greater than about 0.25-3 or 0.5 – 3.0g/ days (such as 0.5,0.75,1,1.25,1.5,1.75,2,2.5,3 or more g/ days).The daily dose of HMB can be approximately, be less than about or be greater than about 0.20 – 3.0g/ days (such as 0.2,0.4,0.5,0.75,1,1.5,2,2.5,3 or more g/ days).The daily dose of KIC can be approximately, be less than about or be greater than about 0.2 – 3.0g/ days (such as 0.2,0.4,0.5,0.75,1,1.25,1.5,1.75,2,2.5,3 or more g/ days).

The dosage of leucine or its metabolite can be therapeutic dose.The dosage of leucine or its metabolite can be sub-therapeutic dose.Leucic sub-therapeutic dose can be approximately, be less than about or be greater than about 0.25 – 3.0g (such as 0.25,0.5,0.75,1,1.25,1.5,1.75,2,2.5,3 or more g).Leucic sub-therapeutic dose can be approximately, be less than about or be greater than about 0.25 – 3.0g/ days (such as 0.25,0.5,0.75,1,1.25,1.5,1.75,2,2.5,3 or more g/ days).In some embodiments, said composition comprises the leucine being less than 3.0g daily dose.The sub-therapeutic dose of HMB can be approximately, be less than about or be greater than about 0.05 – 3.0g (such as 0.05,0.1,0.2,0.4,0.5,0.75,1.5,2,2.5,3 or more g).The sub-therapeutic dose of HMB can be approximately, be less than about or be greater than about 0.05 – 3.0g/ days (such as 0.05,0.1,0.2,0.4,0.5,0.75,1.5,2,2.5,3 or more g/ days).The sub-therapeutic dose of KIC can be approximately, be less than about or be greater than about 0.1 – 3.0g (such as 0.1,0.2,0.4,0.5,0.75,1,1.25,1.5,1.75,2,2.5,3 or more g).The sub-therapeutic dose of KIC can be approximately, be less than about or be greater than about 0.1 – 3.0g/ days (such as 0.1,0.2,0.4,0.5,0.75,1,1.25,1.5,1.75,2,2.5,3 or more g/ days).

Can be that the dosage of unit dose can comprise nicotinic acid, nicotinamide riboside or nicotinic acid metabolite, its can be approximately, more than about or be less than about 0.01,0.05,0.1,0.5,1,2,5,10,20,40,60,80,100,200,250,400,500,800,1000 or the nicotinic acid of 1500mg, nicotinamide riboside or nicotinic acid metabolite.Said composition can comprise about 1-100,5-50 or 10-20mg nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite.In some embodiments, unit dose can comprise at least about the nicotinic acid of 1mg and/or nicotinamide riboside and/or nicotinic acid metabolite.In some embodiments, unit dose can comprise the nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite that are less than 250mg.This dosage can for expecting that the experimenter used regulates.Such as, the dosage being applicable to dog class may be less than the dosage of applicable people.The amount of nicotinic acid as herein described, nicotinamide riboside and/or nicotinic acid metabolite can every day or simultaneously use.Amount as herein described can use with a dosage or every day with multiple dosage separate administration.

In some embodiments, said composition comprises nicotinic acid and nicotinamide riboside, and the total amount of nicotinic acid and nicotinamide riboside can be approximately, more than about or be less than about 0.01,0.05,0.1,0.5,1,2,5,10,20,40,60,80,100,200,250,400,500,600,800,900,1000 or 1500mg.

In other embodiments, the daily dose of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can be approximately, higher than about or lower than about 0.0001mg/kg (mg nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite/kg accept the experimenter of this dosage), 0.005mg/kg, 0.01mg/kg, 0.5mg/kg, 1mg/kg, 2.5mg/kg, 5mg/kg, 7.5mg/kg, 10mg/kg, 12.5mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 50mg/kg, 75mg/kg, 100mg/kg or higher.

Can be that the dosage of unit dose can comprise approximately, to be less than about or more than the resveratrol of about 1,5,10,25,35,50,51,75,100,150,200,250,300,350,400,450,500 or more mg.Said composition can comprise about 5-500,30-250 or 35-100mg resveratrol.In some embodiments, unit dose can comprise at least about 35mg resveratrol.The amount of resveratrol as herein described can every day or simultaneously use.Amount as herein described can use with a dosage or every day with multiple dosage separate administration.

The low daily dose of resveratrol can comprise approximately, be less than about or be greater than about 0.5mg/kg (mg resveratrol/kg accepts the experimenter of this dosage), 1mg/kg, 2.5mg/kg, 5mg/kg, 7.5mg/kg, 10mg/kg, 12.5mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 50mg/kg or higher; The medium daily dose of resveratrol can comprise approximately, is less than about or is greater than about 20mg/kg, 25mg/kg, 50mg/kg, 75mg/kg, 100mg/kg, 125mg/kg, 150mg/kg, 175mg/kg, 200mg/kg, 250mg/kg or higher; And the high daily dose of resveratrol can comprise approximately, is less than about or is greater than about 150mg/kg, 175mg/kg, 200mg/kg, 225mg/kg, 250mg/kg, 300mg/kg, 350mg/kg, 400mg/kg or higher.Be defined as low, in or high dosage scope can depend on the experimenter accepting this dosage, and different between subjects.

In some embodiments, the compositions that can be formulated as unit dose can comprise (a) at least about 250mg leucine and/or at least about one or more leucine metabolite described in 25mg.Said composition can comprise further at least about 35mg resveratrol.

In some embodiments of the present invention, this coupling compositions can have the designated ratio of leucine and/or its metabolite and nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite.This ratio of specifying can provide the effective and/or collaborative treatment to the hyperlipemia patient's condition, such as, it can be measured as the reduction of total lipid content, the reduction of cholesterol levels, the reduction of triglyceride levels, the reduction of LDL level, the alleviating and/or the rising of HDL level of body weight.The ratio of leucine aminoacid and/or its metabolite and nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can be mass ratio, mol ratio or volume ratio.

In some embodiments, compositions can comprise (a) leucine and/or its metabolite (comprising HMB) and (b) nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, and wherein (a) and the mass ratio of (b) can be approximately, be less than about or be greater than about 0.1,0.5,1,2,5,10,15,25,50,75,100,200,300,350,400,450,500,550,600,650,700,750 or 800.In some embodiments, (a) is at least about 25 with the mass ratio of (b).In some embodiments, (a) is at least about 50 with the mass ratio of (b).Said composition can also comprise the nicotinic acid of minimum and/or nicotinamide riboside and/or nicotinic acid metabolite, as 5,10 or 50mg nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite, or the nicotinic acid of certain limit and/or nicotinamide riboside and/or nicotinic acid metabolite amount, as 5-250mg nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite.

In other embodiments, compositions can comprise (a) nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite and (b) resveratrol, and wherein (a) and the mass ratio of (b) can be approximately, be less than about or be greater than about 0.01,0.05,0.1,0.5,1,2,5,10,20,50,100,200,300,350,400,450,500,550,600 or 650.

In some embodiments, the dosage of leucine, leucic any metabolite, nicotinic acid, nicotinamide riboside, any nicotinic acid metabolite and resveratrol can be designed, to reach appointment physiological concentration or the cyclical level of leucine, leucine metabolite, nicotinic acid, nicotinamide riboside, nicotinic acid metabolite and/or resveratrol.This physiological concentration can be the cyclical level recorded in the serum or blood flow of experimenter.This experimenter can be human or animal.The dosage selected can change as body weight, energy metabolism speed, hereditism, race, height or any other feature according to the feature of experimenter.

In some embodiments, the compositions of selected dosage can be applied to experimenter, make this experimenter reach the required cyclical level of said composition.The required cyclical level of component can be treatment effect level or sub-treatment level.

In unit dose, leucic amount can be such, and it makes leucic cyclical level in experimenter be about or be greater than about 0.25mM, 0.5mM, 0.75mM or 1mM.The administration of about 1,125mg leucine (such as free leucine) can reach the leucine cyclical level of about 0.5mM in experimenter.The administration of about 300mg leucine (such as free leucine) can reach the leucine cyclical level of about 0.25mM in experimenter.

The required cyclical level of said composition can be at least about 0.25,0.5,0.75, the leucine of 1mM or more.The required cyclical level of said composition can be at least about, be less than about or be greater than the leucine metabolite (as HMB) of about 0.1,0.25,0.5,0.75,1,10,20,40,60 μM or more.The required cyclical level of compositions can be at least about 0.25,0.5,0.75, the KIC of 1mM or more.

The required cyclical level of said composition can be at least about, be less than about or be greater than about 0.1,0.25,0.5,0.75,1,10,20,40,60,80,100,120,200,400,500,1000,1500,2000, the nicotinic acid of 2500 or 3000nM or more and/or nicotinamide riboside and/or nicotinic acid metabolite.The treatment effect level of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite can be 44-111 μM, and it is equivalent to about 10-20 μ g/mL.

The required cyclical level of said composition can be at least about, be less than about or be greater than about 40,60,80,100,120,150,200,300,400,800,1600, the resveratrol of 3000 or 5000nM or more.The dosage selected can based on the feature of experimenter as body weight, height, race or hereditism select.

In some embodiments, compositions comprises leucine and nicotinic acid with the amount of the cyclical level effectively reaching about 0.3-1mM leucine and about 1-100nM nicotinic acid in experimenter.

About 1,125mg leucic oral administration amount can reach the leucic leucine cyclical level of about 0.5mM in experimenter.The leucic oral administration amount of about 300mg can reach the leucine cyclical level of about 0.25mM in experimenter.

The oral administration amount of about 500mgHMB can reach the HMB cyclical level of about 5 μMs of HMB in experimenter.The oral administration amount of about 100mgHMB can reach the HMB cyclical level of about 0.8 μM of HMB in experimenter.

The oral administration amount of about 3,000mg nicotinic acid or nicotinamide riboside can reach nicotinic acid or the nicotinamide riboside cyclical level of about 10 μMs of nicotinic acid or nicotinamide riboside in experimenter.The oral administration amount of about 50mg nicotinic acid or nicotinamide riboside can reach nicotinic acid or the nicotinamide riboside cyclical level of about 10-100nM nicotinic acid or nicotinamide riboside in experimenter.

The oral administration amount of about 1100mg resveratrol can reach the resveratrol cyclical level of about 0.5mM resveratrol in experimenter.The oral administration amount of about 50mg resveratrol can reach the resveratrol cyclical level of about 200nM resveratrol in experimenter.

In some embodiments, can compositions formulated to reach required circulation mol ratio or mass ratio after using one or more compositionss to experimenter.Said composition can be coupling compositions described herein.Mol ratio can be regulated to be beneficial to the bioavailability of one or more components of coupling compositions, picked-up and metabolism processing.Such as, if a kind of bioavailability of component is low, the mole of this component so can be increased relative to other components in coupling compositions.In some embodiments, circulation mol ratio or mass ratio reach in about 0.1,0.5,0.75,1,3,5 or 10,12,24 or 48 hour after application.Circulation mol ratio or mass ratio can remain about or be longer than a period of time of about 0.1,1,2,5,10,12,18,24,36,48,72 or 96 hour.

In some embodiments, the circulation mol ratio of leucine and nicotinic acid or nicotinamide riboside is approximately, is less than about or is greater than about 1,5,10,20,50,100,500,1000,5000 or 10000.In some embodiments, the circulation mol ratio of HMB and nicotinic acid or nicotinamide riboside is for approximately or be greater than about or be less than about 0.01,0.05,0.1,0.5,1,5,10,20,50 or 100.In some embodiments, the circulation mol ratio of nicotinic acid or nicotinamide riboside and resveratrol is approximately, is less than about or is greater than about 0.01,0.05,0.1,0.5,1,5,10,20,50 or 100.

form of medication

Compositions described herein can be mixed with multiple different dosage form.It can orally use as tablet, capsule, pill, granule, Emulsion, gel, multiple beadlet, powder, suspension, liquid, semiliquid, semisolid, syrup, serosity, chewable form, caplet, Perle, lozenge or the solution be encapsulated in capsule.Or said composition can be prepared for suction or for intravenous delivery.Said composition also can be mixed with nasal spray maybe when for during solution form for injection.In some embodiments, said composition can be suitable for oral fluid composition.

Preparation can use technology known in the art to be packaged in inhaler for the compositions sucked.Inhaler can be designed to suck distribution 0.25,0.5 or 1 unit dose at every turn.Inhaler can have the cylinder tank holding and prepare for the present composition sucked, and allows the metering valve distributing the preparation of metered amount when each actuating, and permission operates this device and the actuator guided to by the present composition in experimenter's lung or rank mouth.The compositions of preparation can comprise liquefied gas propellant and possible stabilising carriers.This actuator can have the cooperation discharge nozzle being connected to a tank and the dust cap being used for preventing pollution actuator.Once actuating, the present composition can volatilize, and this causes the droplet forming the present composition.This droplet can rapid evaporation, thus produces the granule of micron size, is sucked subsequently by experimenter.Inhaler and for prepare composition for inhalation method at U.S. Patent number 5,069,204,7,870,856 and U. S. application number 2010/0324002 in describe, these patents and application are incorporated to herein with its entirety by reference.

The compositions of the present invention being suitable for oral administration can be rendered as discrete dosage form, as the capsule of the active component separately containing the scheduled volume as powder or granule, cachet or tablet, or liquid or aerosol spray, solution, the suspension in aqueous or non-aqueous liquid, oil in water emulsion or water-in-oil liquid Emulsion, comprise liquid dosage form (such as, suspension or serosity), and oral dosage form (such as, tablet or bulk powder).Peroral dosage form can be mixed with tablet, pill, lozenge, capsule, Emulsion, lipophilic and hydrophilic suspensions, liquid, gel, syrup, serosity, suspension etc., for individuality to be treated or the oral absorption of patient.Such dosage form can be prepared by any compound method.Such as, active component can with the carrier in combination forming one or more neccessary compositions.The capsule being suitable for oral administration comprises sucking fit (push-fit) capsule be made up of gelatin, and the soft seal capsule be made up of gelatin and plasticizer (as glycerol or Sorbitol).Sucking fit capsule can comprise the active component mixed with filler (such as lactose), binding agent (as starch) and/or lubricant (as Talcum or magnesium stearate) and optional stabilizing agent.Optionally; by compositions is mixed with solid excipient; optionally grind the mixture obtained, and adding suitable auxiliary agent (if needs) post-treatment granulate mixture to obtain tablet or dragee core, obtain for oral compositions of the present invention.Suitable excipient especially filler, as sugar, comprises lactose, sucrose, mannitol or Sorbitol; Cellulose preparation, such as, corn starch, wheaten starch, rice starch, potato starch, gelatin, Tragacanth, methylcellulose, hydroxypropyl emthylcellulose, sodium carboxymethyl cellulose and/or polyvinylpyrrolidone (PVP).Usually, by by active component and liquid-carrier or solid carrier in small, broken bits or both equably and closely mix, then, if needed, be required form by formed product, prepare compositions.Such as, tablet can be prepared by optionally suppressing or be molded together with one or more auxiliary elements.Compressed tablets is prepared by compressing the active component of free-flowing form as powder or granule in suitable machine, this active component optionally with mixed with excipients, this excipient is such as but not limited to binding agent, lubricant, inert diluent and/or surfactant or dispersant.Molded tablet is prepared by being molded the mixture of the powder compound with inert liquid diluent moistening in suitable machine.

Can be incorporated herein disclosed preparation for oral or by inject the liquid form used comprise aqueous solution, suitably seasoned syrup, water or oil suspension and seasoning containing edible oil as the Emulsion of Oleum Gossypii semen, Oleum sesami, Oleum Cocois or Oleum Arachidis hypogaeae semen and elixir and similar pharmaceutical carrier.The natural gum of synthesis is comprised as Tragacanth, arabic gum, alginate, glucosan, sodium carboxymethyl cellulose, methylcellulose, polyvinylpyrrolidone or gelatin for the suitable dispersant of waterborne suspension or suspending agent.

The combined therapy experimenter of Injectable composition and Orally administered composition can be used.

Can adopt the form of such as solution, syrup or suspension for oral liquid preparation, or they can be rendered as dryed product, for rebuilding with water or other suitable carriers before use.This kind of liquid preparation can by conventional means with pharmaceutically acceptable additive as suspending agent (such as, sorbitol syrup, methylcellulose or hydrogenated edible fats), emulsifying agent (as lecithin or arabic gum), non-aqueous carrier (such as, almond oil, grease or ethanol), prepared by antiseptic (such as, methyl parahydroxybenzoate or propyl p-hydroxybenzoate or sorbic acid) and artificial or natural toner and/or sweeting agent.

The preparation of pharmaceutical composition of the present invention (comprise oral and suck preparation) can be carried out according to generally accepted pharmaceutical preparation preparation procedure.See, such as Remington'sPharmaceuticalSciences the 18th edition (1990), E.W.Martin compile, MackPublishingCo., PA.According to desired use and method of application, may need in the preparation of pharmaceutical composition, process magnesium-equilibrium ion compound further.Suitable process can comprise and mix with suitable non-toxic and non-interfering component, sterilizing, be divided into dosage unit and wrap in delivery apparatus.

The present invention comprises anhydrous composition containing active component and dosage form further, because water can promote the degraded of some compounds.Such as, (such as 5%) means as simulation long storage periods that add water can be adopted in this area, to determine the characteristics such as such as shelf life or preparation stability in time.Anhydrous composition of the present invention and dosage form can use anhydrous or low aqueous ingredients to prepare under low moisture or low-moisture conditions.If anticipate in manufacture, packaging and/or storage process and can contact in a large number with moisture and/or humidity, so can the compositions of the present invention containing lactose and dosage form be made anhydrous.Can prepare and store anhydrous composition, its anhydrous nature is kept.Therefore, anhydrous composition can use the known material being exposed to water that can prevent to pack, and they can be included in suitable prescription test kit.Paper tinsel, plastics etc. that the example of suitable packaging includes but not limited to seal, unit-dose container, blister package and strip packaging.

Composition as herein described can conveniently pharmaceutical compounding techniques and pharmaceutical carrier combine in intimately admixing thing.Depend on and use required dosage form, carrier can take various ways.When the compositions for the preparation of peroral dosage form, any conventional drug media all can be used as carrier, such as, water, glycol, oil, alcohol, flavoring agent, antiseptic, coloring agent etc. are adopted when oral liquid (as suspension, solution and elixir) or aerosol; Or the carriers such as such as starch, sugar, microcrystalline Cellulose, diluent, granulating agent, lubricant, binding agent and disintegrating agent can be used when oral solid formulation, do not use lactose in some embodiments.Such as, for solid orally ingestible, suitable carrier comprises powder, capsule and tablet.If needed, tablet can carry out coating by standard aqueous or non-aqueous technology.

Some examples that can serve as the material of pharmaceutically acceptable carrier comprise: (1) sugar, as lactose, dextrose plus saccharose; (2) starch, as corn starch and potato starch; (3) cellulose and its derivates, as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) tragacanth gum powder; (5) Fructus Hordei Germinatus; (6) gelatin; (7) Talcum; (8) excipient, as cocoa butter and suppository wax; (9) oil, as Oleum Arachidis hypogaeae semen, Oleum Gossypii semen, safflower oil, Oleum sesami, olive oil, Semen Maydis oil and soybean oil; (10) dihydroxylic alcohols, as propylene glycol; (11) polyhydric alcohol, as glycerol, Sorbitol, mannitol and Polyethylene Glycol; (12) ester, as ethyl oleate and ethyl laurate; (13) agar; (14) buffer agent, as magnesium hydroxide and aluminium hydroxide; (15) alginic acid; (16) apirogen water; (17) isotonic saline solution; (18) Ringer's mixture; (19) ethanol; (20) phosphate buffered solution; (21) other the nontoxic compatible materials used in pharmaceutical preparation.

The binding agent be applicable in dosage form includes but not limited to that corn starch, potato starch or other starch, gelatin, natural and paragutta are as arabic gum, sodium alginate, alginic acid, other alginate, tragacanth gum powder, guar gum, cellulose and its derivates (such as ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized Starch, hydroxypropyl emthylcellulose, microcrystalline Cellulose, and their mixture.

The lubricant that can be used for being formed compositions of the present invention and dosage form includes but not limited to calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerol, Sorbitol, mannitol, Polyethylene Glycol, other glycol, stearic acid, sodium lauryl sulphate, Pulvis Talci, hydrogenated vegetable oil (such as Oleum Arachidis hypogaeae semen, Oleum Gossypii semen, Oleum Helianthi, Oleum sesami, olive oil, Semen Maydis oil and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, or their mixture.Other lubricant comprises, such as, and the coagulated aerosol of syloid silica gel, synthetic silica or its mixture.Optionally add be less than compositions about 1 % by weight lubricant.

Lubricant can also be combined with tissue barrier, and this tissue barrier includes but not limited to polysaccharide, polysaccharide, biomembrane (seprafilm), interceed and hyaluronic acid.

In disintegrating agent compositions used in the present invention, to provide the tablet that disintegrate occurs when being exposed in aqueous environments.Too many disintegrating agent may produce the tablet of disintegratable in bottle.The generation causing disintegrate may be not enough to very little, and therefore may change speed and degree that active component discharges from dosage form.Therefore, can use neither also not many very little and deleteriously can not change enough disintegrating agents of the release of active component, to form the dosage form of compound disclosed herein.The amount of disintegrating agent used can change according to preparation type and method of application, and those of ordinary skill in the art can easily distinguish.The disintegrating agent of about 0.5% to about 15% (weight) or the disintegrating agent of about 1% to about 5% (weight) can be used in pharmaceutical composition.The disintegrating agent that can be used for being formed compositions of the present invention and dosage form includes but not limited to agar, alginic acid, calcium carbonate, microcrystalline Cellulose, cross-linking sodium carboxymethyl cellulose, crospovidone, polacrilin potassium, sodium starch glycollate, Rhizoma Solani tuber osi or tapioca, other starch, pregelatinized Starch, other starch, clay, other algin, other celluloses, natural gum or its mixture.

Example for the suitable filler of compositions disclosed herein and dosage form includes but not limited to Talcum, calcium carbonate (such as granule or powder), microcrystalline Cellulose, Powderd cellulose, dextrates, Kaolin, mannitol, silicic acid, Sorbitol, starch, pregelatinized Starch, and their mixture.

When expecting to use waterborne suspension and/or elixir to carry out oral, active component wherein can with multiple sweeting agent or flavoring agent, coloring material or dye combinations, and if if required, can combine with emulsifying agent and/or suspending agent and diluent such as water, ethanol, propylene glycol, glycerol and various combination thereof.

Tablet can not coating or by known technology coatings to postpone disintegrate in the gastrointestinal tract and absorption, and within the long term, provide continuous action thus.Such as, time delay material can be adopted as glyceryl monostearate or distearin.The preparation Gong orally using can also be rendered as hard gelatin capsule, wherein active component and inert solid diluent such as calcium carbonate, calcium phosphate or Kaolin mixes, or as Perle, wherein active component and water or oily medium such as Oleum Arachidis hypogaeae semen, liquid paraffin or mixed with olive oil.

In one embodiment, said composition can comprise solubilizing agent to guarantee good dissolving and/or the stripping of compound of the present invention, and the precipitation of compound of the present invention is minimized.This is for the compositions of parenteral use, and such as, the compositions for injecting is particular importance.Also can add solubilizing agent to increase hydrophilic medicament and/or other components as the dissolubility of surfactant, or maintain said composition as stable or homogeneous solution or dispersion liquid.

Said composition can comprise one or more pharmaceutically acceptable additive and excipient further.Such additive and excipient include but not limited to antitack agent, defoamer, buffer agent, polymer, antioxidant, antiseptic, chelating agen, viscosity modifier (viscomodulators), tension regulator (tonicifier), flavoring agent, coloring agent, flavour enhancer, opacifier, suspending agent, binding agent, filler, plasticizer, lubricant and composition thereof.The non-exhaustive listing of the example of excipient comprises monoacylglycerol, magnesium stearate, modified food starch, gelatin, microcrystalline Cellulose, glycerol, stearic acid, silicon dioxide, yellow beeswax, lecithin, hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose and crospovidone.

Compositions described herein also can be mixed with and extend release, sustained release or time controlled released, discharges in time to make one or more components.Delayed release realizes by these one or more components being formulated in the substrate of various material or by microencapsulation.Said composition can be formulated as and discharges one or more components through the time period of 1,4,6,8,12,16,20,24,36 or 48 hour.The release of one or more components can be carried out with speed that is constant or change.

In some embodiments, the present composition as herein described can be mixed with matrix granule agent (matrixpellet), wherein the embed particles of the present composition is in the substrate of water-insoluble plastics, and wrapped up by the film of the water-insoluble plastics containing the lactose granule embedded, produce and maintain the blood plasma level of the present composition in target therapeutic range.In other embodiments, the present composition can be mixed with the continuous release tablet obtained in the following manner: on the core granule formed primarily of the present composition, apply one deck be made up of hydrophobic material and plastics excipient and the coating membrane optionally containing enteric polymer material, to form coated granule, then coated granule and disintegrate excipient are compressed together.Extended release preparation at U.S. Patent number 4,803,080 and 6,426, describe in 091, it is incorporated to herein with its entirety by reference.

Use Co ntrolled release dosage form provided herein, one or more cofactors can than slower speed release viewed in the immediate release formulation that group component is identical in its dosage form.In some embodiments, about 80%, 70%, 60%, 50%, 40%, 30%, 20% or 10% of immediate release formulation speed is less than for Co ntrolled release preparation according to the change speed from the biological sample measured by the concentration change in the stipulated time section being applied to its Cmax.In addition, in some embodiments, concentration change speed is in time less than about 80%, 70%, 60%, 50%, 40%, 30%, 20% or 10% of immediate release formulation speed.

In some embodiments, the time by reaching Cmax with relatively proportional mode increase reduces concentration change speed in time.Such as, the time reaching Cmax increases by 2 times can make concentration change rate reduction about 2 times.Therefore, one or more cofactors can be provided thus make it reach its Cmax with the remarkable speed lower than immediate release formulation.Compositions of the present invention can be formulated as to 24 hours, 16 hours, 8 hours, 4 hours, 2 hours or at least 1 little change that Cmax is provided constantly.It can be about 0.05,0.10,0.25,0.5 or at least 0.8 times that concentration changes the relevant minimizing of speed.In certain embodiments, this is realized by these one or more cofactors being less than about 30%, 50%, 75%, 90% or 95% to blood circulation release in 1 after this using hour.

Optionally, the blood plasma concentration curve that Co ntrolled release formulations display goes out has initial (after such as, using, 2 is little of using latter 4 hours) slope of 75%, 50%, 40%, 30%, 20% or 10% of the immediate release formulation of the identical cofactor be less than containing same dose.

In some embodiments, at initial 1,2,4,6,8,10 or 12 hour, the cofactor rate of release measured in stripping research was less than about 80%, 70%, 60%, 50%, 40%, 30%, 20% or 10% of the immediate release formulation containing identical cofactor.

Co ntrolled release preparation provided herein can adopt various ways.In some embodiments, said preparation is peroral dosage form, comprises liquid dosage form (such as, suspension or serosity) and oral dosage form (such as, tablet or bulk powder), such as but not limited to those dosage forms described herein.

The Co ntrolled release tablet of preparation disclosed herein can be substrate, reservoir (reservoir) or osmosis system.Although three systems are all applicable, latter two systems can possess the ability of the relatively large quality of more excellent encapsulating, and such as, for holding a large amount of single cofactors or holding multiple cofactor, it depends on individual Gene effect.In some embodiments, slow releasing tablet is based on containment system, and the core wherein comprising one or more cofactors is wrapped up by perforated membrane coating, and this coating allows these one or more cofactors to diffuse through when aquation.Due to the combination quality normally gram number of stages of effective ingredient, so effective delivery system can provide optimum result.

Therefore, tablet or pill also can by coating or otherwise compounds, to provide the dosage form of the advantage with prolongation effect.Such as, tablet or pill can comprise interior dosage and external dose component, and the latter is the form of the bag quilt on the former.These two kinds of components can be separated by enteric layer, and enteric layer can be resisted disintegrate under one's belt and allow internal composition intactly to enter duodenum or delayed release.Multiple material can be used for such enteric layer or coating, and these materials comprise the mixture of multiple polymeric acid and polymeric acid and the such as material that lac, spermol and cellulose acetate are such.In some embodiments, the preparation containing multiple cofactor can have with different rates or the different auxiliary factor in different time release.Such as, the extra cofactor layer being mingled with enteric layer can be there is.

The method preparing continuous release tablet is known in the art, for example, see U.S. Patent Publication 2006/051416 and 2007/0065512 or other lists of references disclosed herein.Such as at U.S. Patent number 4,606,909,4,769,027,4,897,268 and 5,395, the method described in 626 can be used for the extended release preparation preparing one or more cofactors determined by the Gene effect of individuality.In some embodiments, use such as at U.S. Patent number 6,919,373,6,923,800,6,929,803 and 6,939, describe in 556 technology prepares preparation.Such as at U.S. Patent number 6,797,283,6,764,697 and 6,635, the additive method described in 268 also can be used for preparing preparation disclosed herein.

In some embodiments, described compositions can be formulated in food composition.Such as, said composition can be beverage or other liquid, solid food, semi-solid food products, and it contains or does not contain food carrier.Such as, said composition can comprise the black tea being supplemented with arbitrary composition described herein.Said composition can be the milk product being supplemented with arbitrary composition described herein.In some embodiments, said composition can be formulated in food composition.Such as, said composition can comprise beverage, solid food, semi-solid food products or food carrier.

In some embodiments, liquid food carrier can be used, as the form as beverage, as complementarity fruit juice, coffee, tea, soda pop, add taste water etc.Such as, this beverage can comprise preparation and liquid component, as various deodorizer or the natural carbohydrate that is present in conventional beverage.The example of natural carbohydrate includes but not limited to: monosaccharide, as glucose and fructose; Disaccharide, as maltose and sucrose; Conventional sugar, as dextrin and cyclodextrin; And sugar alcohol, as xylitol and erythritol.Also natural deodorant can be used as Talin, Stevia rebaudiana (Bertoni) Hemsl extract, Lai Baodi glycosides A (levaudiosideA), liquirtin, and the deodorizer of synthesis is as glucide and aspartame.Also such as flavoring agent, coloring agent and other reagent can be used.Such as, pectic acid and salt, alginic acid and salt thereof, organic acid, protective colloid binding agent, pH adjusting agent, stabilizing agent, antiseptic, glycerol, alcohol or carburization agent can also be used.Also fruits and vegetables can be used herein in the Foods or drinks of preparation containing the preparation discussed.

Alternatively, described compositions can be the snack rod being supplemented with any compositions as herein described.Such as, this snack rod can be chocolate bars, Herba bromi japonici (granola) rod or assorted assorted fruit (trailmix) rod.In another embodiment, dietary supplement of the present invention or food composition are formulated into and have suitable mouthfeel, quality and viscosity with expecting for consumption.Any suitable food carrier can use in food composition of the present invention.Food carrier of the present invention comprises almost any food.The example of this kind of food carrier includes but not limited to food stick (Herba bromi japonici rod, protein rod, candy bar etc.), cereal products (oatmeal, breakfast oatmeal, Herba bromi japonici etc.), bakery (bread, doughnut, cookies, bagel, cake, cake etc.), beverage (milk is main beverage, sports drink, fruit juice, alcoholic beverage, bottled water), wheaten food, corn (rice, Semen Maydis, Herba bromi japonici, rye (Secale cereale L.), Semen Tritici aestivi, flour etc.), egg products, snacks (confection, potato chips, chewing gum, chocolate etc.), meat, fruits and vegetables.In one embodiment, the food carrier adopted herein can cover bad taste (such as bitterness).When needed, proposed herein food composition shows more preferably quality and fragrance than any component described herein.Such as, liquid food carrier can be used to obtain the drink form of food composition of the present invention, as complementarity fruit juice, coffee, tea etc. according to the present invention.In other embodiments, can food carrier used according to the invention obtain generation meal form food composition of the present invention, as complementarity snack rod, wheaten food, bread etc.In other embodiments, chewing gum, the form such as chewable candies or snacks of food composition of the present invention can be obtained by semi-solid foodstuff carrier used according to the invention.

The administration of coupling compositions can daily approximately, to be less than about or more than about 1,2,3,4,5,6,7,8,9,10 times or more times.Experimenter can approximately, be less than about or be greater than about 1,2,3,4,5,6, accept administration in 7,8,9,10,11,12,13,14 or more sky, week or the moons.Unit dose can be the mark of daily dose, and such as daily dose is divided by unit dose number to be administered every day.Unit dose can be the mark of daily dose, and it is daily dose divided by unit dose number to be administered every day, then divided by the unit dose number used (such as sheet number) at every turn.The unit dose number at every turn used can for approximately, to be less than about or more than about 1,2,3,4,5,6,7,8,9,10 or more.The dosage number of every day can for approximately, to be less than about or more than about 1,2,3,4,5,6,7,8,9,10 or more.The unit dose number of every day can by determining divided by unit dose with daily dose, and its can for approximately, to be less than about or more than about 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,6,17,18,19,20 or more unit dose every days.Such as, unit dose can be about 1/2,1/3,1/4,1/5,1/6,1/7,1/8,1/9,1/10.Unit dose can be every feed ration about three/in the lump to experimenter daily three times.Unit dose can be every feed ration about two/in the lump to experimenter daily twice.Unit dose can be every feed ration about four/in the lump to experimenter daily twice, each two unit dose.In some embodiments, unit dose comprises approximately, to be less than about or more than about 50mg resveratrol.In some embodiments, unit dose comprises approximately, to be less than about or more than about 550mg leucine.In some embodiments, unit dose comprises approximately, to be less than about or more than one or more leucine metabolite of about 200mg.

In some embodiments, unit dose (such as comprising one or more leucine metabolite as the unit dose of HMB) daily twice, often next unit dose.Unit dose can comprise more than one capsule, tablet, bottle or entity.

Compositions disclosed herein can comprise fumet further, and can be solid, liquid, gel or Emulsion.

When the present composition used comprises one or more therapeutic agents further and this therapeutic agent had than leucine and/or leucine metabolite or nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite shorter half-life, the unit dosage form of this therapeutic agent and leucine and/or leucine metabolite or nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite correspondingly can be regulated.

method

Compositions of the present invention especially can be used for improving the hyperlipemia patient's condition.In one embodiment, the invention provides and reduce total lipid content or reduce T-CHOL, LDL or triglyceride levels, raising HDL level or reduce the method for atheromatous plaque size, it comprises uses any compositions of the present invention to experimenter in need.Level described herein or content can be the circulation compositions in serum or blood flow, or the total amount in subject.In some embodiments, compositions of the present invention can be used for increasing losing weight of experimenter, and the sirt1 increasing experimenter activates or fat oxidation.In multiple embodiment of the present invention, use compositions with the amount of the leucine and/or its metabolite, nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite and/or resveratrol of sending the collaborative amount of the hyperlipemia patient's condition being enough to improve experimenter to experimenter.In some embodiments, if nicotinic acid, nicotinamide riboside or nicotinic acid metabolite be not having to be applied to experimenter with its therapeutic dose in leucine or leucine metabolite situation, then side effect (such as, cutaneous vasodilation) can be induced.Method described herein can also be used for improving side effect, and can not lose the therapeutic effect of nicotinic acid, nicotinamide riboside or nicotinic acid metabolite.There is provided herein the description to various aspects, feature, embodiment and example.

Comprise to use to nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite the method for the present invention of leucine and/or leucine metabolite go for suffering from hyperlipidemia, be in suffer from hyperlipemia risk in and/or suffer from the experimenter of the patient's condition relevant with hyperlipemia as cardiovascular diseases's condition and use.In some cases, can use to experimenter together with any compositions of the present invention in the additional therapeutic agent of the known effective dose of medical domain or forms of pharmacologically active agents (such as, hyperlipidemia agent).

The feature of hyperlipemia can be high-caliber total lipid content in experimenter or level.The feature of hyperlipemia also can be high-caliber body weight or the BMI of experimenter.The type of lipid can comprise cholesterol, cholesteryl ester, phospholipid and triglyceride.The content of lipid or level can be the cyclical levels measured in the blood flow of experimenter, blood plasma or serum.The content of lipid can also be relevant to the body weight of experimenter.These lipids can be transported as large lipoprotein in blood, based on their density, comprise Chylomicron, very low density lipoprotein (VLDL) (VLDL), intermediate density lipoprotein (IDL) (IDL), low density lipoprotein, LDL (LDL) and high density lipoprotein (HDL).Most of triglyceride can be transported in Chylomicron or VLDL, and most of cholesterol can carry in LDL and HDL.High-level lipid in circulation can cause the lipid accumulation on arterial wall, and causes atheromatous plaque to be formed further, thus makes stricture of artery.The experimenter of suffering from hyperlipidemia can be in the excessive risk suffering from the cardiovascular patient's condition.The feature of hyperlipemia also can be more high-caliber lipoproteins or low-level HDL.The patient's condition that is that experimenter suffers from or that have a risk can be the patient's condition relevant to the lipoprotein of experimenter or the abnormal level of lipid.Compositions of the present invention can be used for changing the level of one or more lipids in experimenter or lipoprotein.In some embodiments, the type of the lipid that its level can affect by the compositions and methods of the invention or lipoprotein can be one or more lipoproteins and/or lipid, includes but not limited to: T-CHOL, triglyceride, HDL, IDL, VLDL or LDL.

Many methods can be used to assess the lipoprotein in experimenter and/or lipid level.These methods may be different from each other on sample type and analytical technology used.The sample type that can be used to measure this kind of level includes but not limited to: serum, blood plasma, whole blood, erythrocyte or tissue sample.When desired, the level of lipoprotein and/or lipid can be measured under empty stomach condition, such as, does not take food at least about 8 hours, 10 hours, 12 hours, 15 hours, 24 hours or even more of a specified duration.

The size of atheromatous plaque or pathological changes can be measured by any method known in the art.Such as, the people such as PhanBA, " Effectsofniacinonglucoselevels, coronarystenosisprogression, andclinicaleventsinsubjectswithnormalbaselineglucoseleve ls (100mg/dl): acombinedanalysisoftheFamilialAtherosclerosisTreatmentSt udy (FATS), HDL-AtherosclerosisTreatmentStudy (HATS), ArmedForcesRegressionStudy (AFREGS), andCarotidPlaqueCompositionbyMRIduringlipid-lowering (CPC) study ", AmJCardiol.2013 February 1, 111 (3): 352-5, with people such as LehmanSJ, " AssessmentofCoronaryPlaqueProgressioninCoronaryCTAngiogr aphyUsingaSemi-QuantitativeScore ", JACCCardiovascImaging.2009 November, the method described in 2 (11): 1262 – 1270.Being used for the limiting examples of the method measuring atheromatous plaque or pathological changes size can be Quantitative Coronary Angiography art.

In some embodiments, if be applied to experimenter individually and when not having leucine, leucine metabolite or resveratrol, the amount of the nicotinic acid in described compositions, nicotinamide riboside and/or nicotinic acid metabolite will not have therapeutic effect to experimenter.In addition, if be applied to experimenter when not having nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite, the amount of leucine, leucine metabolite or resveratrol may not have therapeutic effect to experimenter.But, when nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite are used together with leucine, leucine metabolite or resveratrol, can be observed therapeutic effect." therapeutic effect " described herein be reduce in used experimenter total lipid content, the total cholesterol level of reduction, the triglyceride levels of reduction, the HDL level of raising, the LDL level of reduction or minimizing atheromatous plaque.Therefore, the invention provides a kind of method using compositions, said composition comprises the nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite that (a) leucine and/or one or more its metabolite and (b) exist with sub-therapeutic dose, wherein compared with the component (b) when being used alone, said composition strengthens the treatment to the hyperlipemia patient's condition effectively.Leucic amount in said composition also can be sub-therapeutic dose.

The quantification of therapeutic effect can show, the effect comprising the compositions of the nicotinic acid of (a) leucine or leucine metabolite and (b) sub-therapeutic dose, nicotinamide riboside or nicotinic acid metabolite is greater than hypothesis (a) and (b) effect using separately (a) or (b) for predicting during simple synergistic effect, and therefore this effect is synergitic.Cooperative effect can be quantified as the effect measured of the prediction simple superposition effect higher than said composition component.Such as, as the effect used separately relative to contrast generation 10% of fruit component (a), the effect used separately relative to contrast generation 15% of component (b), and comprising the effect used relative to contrast generation 60% of the compositions of (a) and (b), then cooperative effect will be 60%-(15%+10%) or 35%.

In some embodiments, the nicotinic acid of therapeutic dose, nicotinamide riboside and/or nicotinic acid metabolite can cause side effect, and the feature of this side effect can be the increase of cutaneous vasodilation.The increase of cutaneous vasodilation can be significant clinically.Compared with the nicotinic acid of therapeutic dose, nicotinamide riboside and/or nicotinic acid metabolite, the nicotinic acid of sub-therapeutic dose, nicotinamide riboside and/or nicotinic acid metabolite can not cause significant cutaneous vasodilation clinically, or can reduce the degree of cutaneous vasodilation in used experimenter.Described herein compositions and method comprise the nicotinic acid of the sub-therapeutic dose that will use together with leucine and/or leucine metabolite, nicotinamide riboside and/or nicotinic acid metabolite, to make the nicotinic acid of sub-therapeutic dose, nicotinamide riboside and/or nicotinic acid metabolite obtain medical treatment the effect of degree, and can not cause usually can by the side effect of the nicotinic acid of therapeutic dose, nicotinamide riboside and/or the nicotinic acid metabolite degree caused when not using together with leucine and/or leucine metabolite.The level of cutaneous vasodilation can be measured by any method that medical domain is known, as comprised the method for Laser-Doppler effusion meter.In identical therapeutic effect level (such as, cholesterol levels is made to reduce at least 5%) under, compared with the nicotinic acid not containing leucine and/or leucine metabolite, nicotinamide riboside and/or nicotinic acid metabolite, the level of the cutaneous vasodilation caused by compositions of the present invention can be lower.Such as, lower than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80% or 90% of the level caused by the nicotinic acid of therapeutic dose, nicotinamide riboside and/or nicotinic acid metabolite.

The amount of leucine and/or leucine metabolite can be at least about 25,50,75,100,150,200,250,300,350,400,450,500mg.The sub-therapeutic dose of nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite can be less than 1g, 500,250,100,50 or 10mg.The amount of nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite can be about 1-100mg.The amount of nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite can reach about 1-100nM, higher than about 100nM or the cyclical level at least about the nicotinic acid of 10nM, nicotinamide riboside and/or nicotinic acid metabolite.

Therefore, multi-component combination described herein (as nicotinic acid/leucine, nicotinic acid/leucine/resveratrol, nicotinamide riboside/leucine, and nicotinamide riboside/leucine/resveratrol) useful or collaborative effect can be had to reducing total lipid content, reducing total cholesterol level, reduce triglyceride levels, improve HDL level and/or reducing LDL level.In some embodiments, compared with the initial level of the lipoprotein before being applied to experimenter and/or lipid, compositions described herein and method can make the level of the lipoprotein in experimenter and/or lipid change at least about 1% effectively, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% or 60% or even more.This level can reduce about 19%-24%, 14%-29%, 12%-35%, 10-40%, 8%-45%, 5%-50%, 2%-60% or 1%-70%.This level can be cyclical level.

In some embodiments, compared with the initial size of the atheromatous plaque before being applied to experimenter, compositions described herein and method can make the atheromatous plaque size in experimenter reduce at least about 1% effectively, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% or 60% or even more.This level can reduce about 19%-24%, 14%-29%, 12%-35%, 10-40%, 8%-45%, 5%-50%, 2%-60% or 1%-70%.

It may be useful that being applied in of the compositions of increase SIRT1 and SIRT3 activity disclosed herein needs in any experimenter of metabolic activation hepatocyte, adipose cell or its one or more muscle (such as, skeletal muscle, smooth muscle or cardiac muscle or its flesh) cell.Experimenter suffers from cachexia or amyotrophic experimenter.Increase SIRT3 activity also can be used for increasing or keep the body temperature of such as low temperature experimenter, and increase SIRT1 activity and be conducive to treatment hyperlipemia, diabetes (type 2 diabetes mellitus) and glucose tolerance and reduce and reduce the inflammatory response of experimenter.The increase of the metabolic activation of hepatocyte, adipose cell or its one or more muscle may be used for reducing lipid content and increasing losing weight of experimenter.The content of lipid and lipoprotein or level can reduce.

Increase SIRT3 activity and also can be used for treatment or prevention hyperlipemia, cardiovascular disease, reduced blood pressure by vasodilation, increase cardiovascular health, and increase the contractile function (such as, by affecting smooth muscle) of vascular tissue such as blood vessel and tremulous pulse.Usually, the activation of SIRT3 can be used to the metabolism of the muscle of cell cultured supernatant, adipose cell or any type (such as, the muscle of intestinal or digestive system or urinary tract), and can be used to thus control intestinal movement, such as constipation and incontinence.It also may be useful that SIRT3 activates in erection disturbance.It can also be used to stimulate motility of sperm, such as, be used as fertility drug.Other embodiments that wherein increase SIRT3 will be useful comprise muscle transformation (after operation or accident), the increase of muscle quantities and the increase of athletic performance.

Therefore, the invention provides the method producing beneficial effect by taking in nicotinic acid and/or nicotinamide riboside and/or its any metabolite and the protein of SIRT1 or SIRT3 or the leucine of activity level and/or leucine metabolite can be improved.The activity of SIRT1 and SIRT3 in the myocyte of experimenter and/or hepatocyte can improve.These methods promote effectively, increase or stimulate following one or more: the calorie restriction in simulation hepatocyte or myocyte or the benefit of taking exercise, increase mitochondrion biology to occur or metabolism, improve the mitochondria activity in hepatocyte or myocyte and/or persistency, make myocyte to insulin sensitivity, increase the fatty acid oxidation in myocyte, reduce the active oxygen (ROS) in myocyte, increase PGC-1 α and/or UCP3 in hepatocyte or myocyte and/or GLUT4 expression, and activate the protein kinase (AMPK) of the AMP activation in hepatocyte or myocyte.Various types of myocyte can be contacted according to the present invention.In some embodiments, this myocyte is Skeletal Muscle Cell.In certain embodiments, this myocyte is slow constrictor cell, as musculus soleus cell.

Can by oral or by other any methods, compositions is applied to experimenter.Oral administration methods comprises to be used as liquid, solid or semisolid compositions, and it can adopt the form of dietary supplement or food.

Said composition can regularly be used.Such as, said composition can daily 1,2,3,4 time or frequently.Can use experimenter every 1,2,3,4,5,6 or 7 day.In some embodiments, said composition daily 3 times.Use can with meal time of experimenter simultaneously.The cycle that treatment or diet supplement can be about 1,2,3,4,5,6,7,8 or 9 day, 2 weeks, 1-11 individual month or 1 year, 2 years, 3 years, 4 years, 5 years or even more of a specified duration.In some embodiments of the present invention, the dosage used to experimenter in treatment cycle can change or remain unchanged.Such as, can increase in administration period or reduce and daily measure.

The length of time of application section and/or dosage can be determined by the clinician of doctor or any other type.Doctor or clinician can observe the reaction of experimenter to the compositions used, and regulate administration based on the performance of experimenter.Such as, dosage can be increased to the experimenter showing energy adjustment decreased effectiveness, to reach the result of expectation.

In some embodiments, the component in described compositions can be used by identical approach simultaneously together, or separate administration.Component in said composition also can be used subsequently.In some embodiments, the leucine in said composition and/or leucine metabolite can be used to experimenter together with nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite.In some embodiments, the component in said composition can be used by identical or different route of administration.Such as, leucine and/or leucine metabolite can be Orally administered, and nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite can be used via intravenous injection.Each in metabolite can be used via identical or different route of administration.

In some embodiments, can optimize for given experimenter the compositions that this experimenter is used.Such as, the specific components in the ratio of leucine and/or leucine metabolite and nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite or coupling compositions can be regulated.Different leucines can be had and/or leucine metabolite is assessed experimenter from after the ratio of nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite or one or more compositionss of different coupling composition component using experimenter, then select this ratio and/or specific components.

Another aspect provides the effect obtaining expectation after using the time period that coupling compositions described herein specifies in one or more experimenter.Such as, said composition is being used to experimenter and can observe after 1,2,3,4,6,8,10,12,24 or 52 weeks the beneficial effect of compositions as herein described.

The invention provides a kind of method for the treatment of experimenter, it comprises a collection of experimenter differentiating to be applicable to treatment.This discriminating step can comprise one or more examinations test or analyze.Such as, experimenter that is that be identified as hyperlipemia or that have Body Mass Index (BMI) and/or the body weight exceeding meansigma methods or be significantly greater than meansigma methods can be selected to treat.This experimenter can be overweight or fat, and this can exceed ideal value by the body weight of experimenter or BMI indicates higher than 25,30,40 or 50.The body weight of this experimenter can exceed about 50,75,100,125,150,160,170,180,190,200,210,220,230,240,250,260,270,280,290,300,310,320,330,340,350,360,370,380,390 or 400 pounds.Also the experimenter of high fat diet can be selected to be used for the treatment of.Then can treat the experimenter differentiated by one or more compositionss described herein.Such as, by the coupling compositions comprising nicotinic acid and branched-chain amino acid, it can be treated.

Present invention also offers the method preparing compositions described herein.In some embodiments, the preparation of compositions described herein comprises mixing or combines two or more components.These components can comprise nicotinic acid, nicotinamide riboside and/or nicotinic acid metabolite and leucine or its metabolite (as HMB or KIC).The amount of component or ratio can be amount as herein described or ratio.Such as, the mass ratio of leucine compared with resveratrol can be greater than about 80.

In some embodiments, described compositions with pharmaceutical active or therapeutic agent, carrier and/or excipient composition or can also mix.This document describes the example of this type of component.Coupling compositions can be formulated as the unit dose such as such as tablet, capsule, gel capsule, slow releasing tablet.

In some embodiments, preparation compositions is to obtain the solid composite comprising the mixture of the homogenizing substantially of one or more components, thus these one or more components are dispersed in said composition, make it possible to said composition is easily subdivided into same effective unit dosage forms, as tablet, pill and capsule.

test kit

Present invention also offers test kit.This test kit comprises one or more compositionss as herein described be in suitable packaging, and may further include written material, and this written material can comprise the discussion, side effect list etc. of operation instruction, clinical research.Such test kit can also comprise the information such as such as scientific references, package insert material, clinical test results and/or these summary, these information show or establish activity and/or the advantage of said composition, and/or describe administration, use, side effect, drug interaction or other information useful to health care provider.Such information based on the result of multiple research, such as, can use the result relating to the research of the laboratory animal of In vivo model and the research based on human clinical trial.Test kit can comprise one or more unit dose described herein.In some embodiments, test kit comprise approximately, be less than about or more than about 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,30,31,60,90,120,150,180,210 or more unit dose.Operation instructions can comprise administration and illustrate, as the daily explanation of 1,2,3,4,5,6,7,8,9,10 time or more time.Such as, test kit can comprise the unit dose as tablet supply, and each tablet is packaging separately, according to the number of unit dose when using at every turn, multiple tablet is packaging (such as paired tablet) separately, or all tablets (such as in bottle) packaging together.As another example, test kit can comprise the unit dose as bottled drink supply, and this test kit comprises 1,2,3,4,5,6,7,8,9,10,11,12,13,14,24,28,36,48,72 or more bottle.

This test kit can comprise another kind of medicament further.In some embodiments, compound of the present invention and medicament provide as the independent compositions in container independent in test kit or pack.In some embodiments, compound of the present invention and medicament provide as the single compositions in the container of test kit or pack.Suitable packaging and additive products (such as, for liquid preparation measuring cup, make the minimized Foilpac of air exposure etc.) be known in the art and can be included in test kit.Test kit as herein described can be provided, sells and/or promotes to health care provider, comprise doctor, nurse, pharmacists, prescription official etc.In some embodiments, test kit also can direct marketing to consumer.

In some embodiments, test kit can comprise supplies in many days of unit dose.This unit dose can be any unit dose as herein described.This test kit can comprise the description of the supply in many days instructing administration of unit doses within a period of time of many days.Supply in these many days can be a supply in month, supply in 30 days or many week supplies.Supply in these many days can be 90 days, 180 days, 3 months or supply in 6 months.This test kit can comprise unit dose every day of packaging, such as the packaging of 1,2,3,4 or 5 unit dose.This test kit can substitute bar with other dietary supplement, vitamin and meals, mixture is packed together with beverage.

embodiment

embodiment 1: nicotinic acid/leucine and nicotinic acid/leucine/resveratrol are in vitro in myocyte the impact of Sirt1 level

Have studied the purposes comprising nicotinic acid and leucic compositions as described herein, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.Said composition activates the Sirt1 in myocyte and can be used to improve the hyperlipemia patient's condition.Be investigated the compositions comprising resveratrol further.

By C2C12 mouse muscle-forming cell (American type culture collection) with 8000 cell/cm ²(10cm ²culture dish) density inoculation, and to grow under 37 DEG C with 5%CO2 improveing in Eagle culture medium (DMEM) (growth medium) containing 10% hyclone (FBS) and antibiotic Dulbecco.In order to carry out the differentiation of C2C12 cell, Growth of Cells to 100% is converged, transfer to division culture medium (DMEM containing 2% horse serum and 1% Pen .-Strep), and supplement fresh division culture medium every day until myotube is fully formed (3 days).

In order to find the sub-therapeutic dose studied variable not being had to influential nicotinic acid, carry out dose-response research by the nicotinic acid using variable concentrations to cell.Finding that the Nicotinic of independent <100nM does not affect, therefore experimental concentration is set in that this is below horizontal, is 10nM.Then with leucine and HMB combined test this sub-treatment level of nicotinic acid.Under leucine and HMB are in certain concentration, previously showed that this concentration can reach in diet or fill-in, and separately therapeutic effect (leucine is 0.5mM, and HMB is 5 μMs) there is no to these variablees when using separately.

10nM nicotinic acid (NA) is used to C2C12 cell myotube, 10nM nicotinic acid and 0.5nM leucine (NA/Leu), 10nM nicotinic acid and 0.5nM leucine and 200nM resveratrol (NA/R/Leu), 200nM resveratrol and 0.5nM leucine (R/Leu), and 10 μMs of nicotinic acid 24 hours.

Western blot analysis is carried out with the SIRT1 antibody available from CellSignaling (Danvers, MA).Protein is measured by BCA test kit (ThermoScientific).Altogether 35 μ g from the protein of cell lysate at 10%Tris/HCL polyacrylamide gel (standard precast gel, Bio-RadLaboratroies, Hercules, CA) upper parsing, be transferred to pvdf membrane, incubation in the Block buffer (TBS containing 3%BSA), with first antibody incubation, washing and with the second antibody incubation of horseradish peroxidase.BioRadChemiDoc instrument and software (Bio-RadLaboratories, Hercules, CA) is used to carry out visual and chemiluminescence detection.Use ImageLab4.0 (Bio-RadLaboratories, Hercules, CA) to assess band intensity, wherein correct for background and loading contrast.Sirt1 detects at 104-115kDA place.By one way analysis of variance to data analysis, least significant difference inspection is used to be separated significantly different cell means.

Find that nicotinic acid-leucine stimulates the Sirt1 in C2C12 myotube synergistically, its effect and resveratrol-leucine are quite (Fig. 2, p<0.05).Independent nicotinic acid has no significant effect Sirt1 level.The triple combination of leucine (10nM)/resveratrol (200nM) and nicotinic acid (10nM) has played significantly larger effect, and Sirt1 level improves 200% (p=0.0001).

embodiment 2: nicotinic acid/leucine and nicotinic acid/leucine/resveratrol are in vitro in adipose cell the impact of P-AMPK/AMPK level

Have studied the purposes comprising nicotinic acid and leucic compositions as described herein, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.Said composition improves defying age enzymatic pathway and exports, and comprises AMPK---the signaling molecule in defying age enzymatic pathway, and the p-AMPK/AMPK level in adipose cell, and can be used to improve the hyperlipemia patient's condition.Be investigated the compositions comprising resveratrol further.

By 3T3-L1 PECTORAL LIMB SKELETON (American type culture collection) with 8000 cell/cm ²(10cm ²culture dish) density inoculation, and in 37 DEG C and 5%CO in containing 10% hyclone (FBS) and antibiotic Dulbecco improvement Eagle culture medium (DMEM) (growth medium) ₂lower growth.The 3T3-L1 PECTORAL LIMB SKELETON the converged standard differentiation culture medium be made up of the DMEM culture medium being supplemented with 10%FBS, 250nM dexamethasone, 0.5mM3-isobutyl group-1-methylxanthine (IBMX) and 1% Pen .-Strep is induced, to be divided into adipose cell.PECTORAL LIMB SKELETON maintains 3 days in this division culture medium, is incubated at subsequently in growth medium.Within every 2-3 days, to culture again feed supplement, reach before carrying out chemical treatment to make the cell of >90% and break up completely.

3T3-L1 cell to differentiation uses 10nM nicotinic acid (NA), 10nM nicotinic acid and 0.5nM leucine (NA/Leu), 10nM nicotinic acid and 0.5nM leucine and 200nM resveratrol (NA/R/Leu), 200nM resveratrol and 0.5nM leucine (R/Leu), and 10 μMs of nicotinic acid 24 hours.

Western blot analysis is carried out with the antibody for AMPK and phosphoric acid-AMPK α (Thr172) available from CellSignaling (Danvers, MA).Protein is measured by BCA test kit (ThermoScientific).Altogether 30 μ g from the protein of cell lysate at 10%Tris/HCL polyacrylamide gel (standard precast gel, Bio-RadLaboratroies, Hercules, CA) upper parsing, be transferred to pvdf membrane, incubation in the Block buffer (TBS containing 3%BSA), with first antibody (P-AMPK) incubation, washing and with the second antibody incubation of horseradish peroxidase.BioRadChemiDoc instrument and software (Bio-RadLaboratories, Hercules, CA) is used to carry out visual and chemiluminescence detection.Use ImageLab4.0 (Bio-RadLaboratories, Hercules, CA) to assess band intensity, wherein correct for background and loading contrast.AMPK detects at 62kDA place, and P-AMPK detects at 64-66kDA place.By one way analysis of variance to data analysis, least significant difference inspection is used to be separated significantly different cell means.

Find that 10nM nicotinic acid activates AMPK when using separately to have no significant effect (Fig. 3).Nicotinic acid-leucic combination have stimulated AMPK significantly and activates, reach the degree (p<0.01) suitable with leucine-resveratrol, as increase by P-AMPK/AMPK prove, and the triple combination of nicotinic acid-leucine-resveratrol is not significantly different from any one (Fig. 2) in two yuan of leucines combinations.

embodiment 3: nicotinic acid/leucine and nicotinic acid/leucine/resveratrol are in vivo to Caenorhabditis elegans in the impact of fat content

Have studied the purposes comprising (a) nicotinic acid and (b) leucic compositions as described herein, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.After using said composition to experimenter, said composition reduces the lipid content in experimenter.

Caenorhabditis elegans (Celegans) anthelmintic (N2Bristol wild type) available from the hidden rhabditida hereditism center (CaenorhabditisGeneticsCenter) (CGC) of University of Minnesota (UniversityofMinnesota), and using escherichia coli (OP50) on the standard NGM plate of food source in 20 DEG C of growths.For process, make ovum incubated overnight on the flat board of hunger.Then synchronized L1 larva is transferred to containing shown handled thing, with escherichia coli raise NGM plate on about 35 hours, with reach L4/ children the manhood.All handled things are all added into agar.Handled thing comprises 10nM nicotinic acid and 0.5mM leucine.

Method as herein described is used to measure fat content, protein content, the fatty acid oxidation of Caenorhabditis elegans anthelmintic.

The oil red (Oil-Red) being used for quantizing fat content is dyeed, washes treated L4/ children adult worms off 3 times from plate with PBS, and be collected in 15ml conical pipe, subsequently with the centrifugal 30sec of 1000g.Abandoning supernatant also uses 10mlPBS washing precipitate.After centrifugal, discard the supernatant except 400 μ l, this 400 μ l is transferred in new 1.5mleppendorf pipe.Then 2xMRWB (160mMKCl, 40mMNaCl, the 14mMNa of 500 μ l is added ₂eGTA, 1mM spermidine HCl, 0.4mM spermine, 30mMNaPIPESpH7.4,0.2% beta-mercaptoethanol) and 20% paraformaldehyde of 100 μ l, sample is at room temperature rocked 60 minutes gently.Then by pipe with the centrifugal 30sec of 1500g, then suction and with PBS washing once, recentrifuge, and suction be 300 μ l.Add 700 μ l isopropyl alcohols, mix by pipe is put upside down, and in incubation at room temperature 15min under shaking gently.To manage centrifugal with after removing isopropyl alcohol, (0.5g oil red O, in 100ml anhydrous isopropyl alcohol, balances 2 days by room temperature stirring, then the ddH of 4 times of volumes to add to anthelmintic the oil red-O-dye solution that 1ml60% filters ₂o mixes with the dye solution of 6 times of volumes, and at room temperature balances 15min, then with the bore filter of 0.2M), and rotate on shaking table and spend the night.By anthelmintic with the centrifugal 30sec of 1200g, use ddH subsequently ₂o washs 4 times to remove any unconjugated stain.In order to quantize, by adding 100% isopropyl alcohol eluting oil red O from cell, and use BiotekSynergyHT microplate reader (BioTek, Winooski, VT, USA) under 540nm wavelength, determine the optical density of 200 μ l equal portions (triplicate/sample).PierceBCA Protein Assay Kit is used data to be normalized protein content.

In order to by western blot method determination protein content, with M9 buffer, treated L4/ children adult worms is washed off from plate, and collect in microcentrifugal tube.After centrifugal (500g, 5min), take out supernatant to about 100 μ l.Then add 250 μ lRIPA buffer and add protease and inhibitors of phosphatases mixture.By sample homogenization, then at 40 DEG C with the centrifugal 10min of 16,000g.The supernatant of clarification is used for further experiment.Use PierceBCA Protein Assay Kit determination protein content.

(BruckbauerA, ZemelMB.Synergisticeffectsofmetformin, resveratrol, andhydroxymethylbutyrateoninsulinsensitivity.Diabetes, MetSyndObesity2013 as previously mentioned; 6:93-102), through slightly revising, measure fatty acid oxidation with XF24 analyser (SeahorseBioscience, Billerica, MA, USA) by the consumption rate measuring cetylate stimulation.With M9 buffer, treated L4/ children adult worms is washed off from plate, and collect in 15ml conical pipe.After centrifugal (1000g, 1min), removing supernatant, and anthelmintic precipitate is diluted to the concentration of 40 anthelmintics/μ l.Anthelmintic is remained in frozen water in plating process and move to limit it, and 5 μ l anthelmintic solution are added to (about 200 anthelmintic/holes) in each hole of Seahorse island, 24-hole plate.Insert dividing plate (Screens), and in each hole, add the M9 buffer that 595 μ l contain shown handled thing.Each plate cools 10min before starting to measure.The temperature of instrument is arranged in experimentation and maintains 29 DEG C.

By one way analysis of variance to data analysis, and least significant difference inspection is used to be separated significantly different cell means.

As shown in Figure 4, we determine the lipid content in Caenorhabditis elegans.Find that Caenorhabditis elegans is exposed to leucine (0.5mM)-nicotinic acid (10nM) and combines and within 24 hours, cause total lipid content reduction by 33% compared with untreated matched group.

embodiment 4: nicotinic acid/leucine and nicotinic acid/leucine/resveratrol in vivo to triglyceride, LDL, the impact of HDL and cholesterol levels and atheromatous plaque size

In order to assess effect of title compound, use the title compound comprising (a) nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite and (b) leucine and/or leucine metabolite as herein described to mice, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.Said composition can be used to the triglyceride, LDL and the cholesterol levels that reduce after using said composition to mice in mice and the size reducing atheromatous plaque.Be investigated the compositions comprising resveratrol further.

Ldl receptor knocks out (LDLRKO) mice is available from JacksonLaboratories (BarHarbor, ME), and grouping is raised under the environment and regular light dark period of room temperature, humid control.Mice is freely obtained before treatment containing 0.2% (weight) cholesterol and the atherogenicity diet 8 weeks from 10% calorie of saturated fat (Petiolus Trachycarpi oil) and water.For process, give 250mg nicotinic acid/kg diet that mice (a) is independent, (b) 250mg nicotinic acid/kg diet and 24g leucine/kg diet, (c) 50mg nicotinic acid/kg diet, 24g leucine/kg diet and 12.5mg resveratrol/kg diet, (d) 24g leucine/kg diet and 12.5mg resveratrol/kg diet, 10g nicotinic acid/kg diet that (e) is independent.Described process continuous administration 60 days.Comprise two matched groups for comparing: negative control group accepts not containing the atherogenicity diet of handled thing, double-negative matched group accepts not containing the normal diet of handled thing.All chemicals are all purchased from Sigma.

The blood sample of all groups of small mouses obtains from the tail of mice all after application on the the 7th, 30 and 60 day.Measure the serum/plasma level of triglyceride, cholesterol, LDL and HDL.Cutaneous vasodilation is by Laser-Doppler flowmeter survey.At the 60th day, put to death mice to quantize Aortic Plaque.In brief, dissect and remove adhere to fatty tissue after, aorta is immersed in above-mentioned buffer and spends the night.After removal adventitia, longitudinally cut by aorta, smooth ground nail is being dissected on wax, and with oil red O stain for micro image analysis.Go out the profile in the aortic disease region of total aorta regions and dyeing with blinded manner hand drawing, and use AdobePhotoshopCS3 to analyze, as accounting for the percentage ratio of total aorta regions to determine pathological changes size.

Result

Compared with the double-negative matched group accepting normal diet, only accept can not show triglyceride, LDL and cholesterol levels significantly higher in blood flow containing the mice of the atherogenicity diet of handled thing.For processed group, group (a) can demonstrate the triglyceride similar with the negative control group only accepting atherogenicity diet, LDL, cholesterol and HDL level, does not have statistically evident difference.Compared with only accepting the negative control group of atherogenicity diet, group (b), (c) and (e) can show triglyceride, LDL and cholesterol levels significantly lower in blood flow, and significantly higher HDL level, and these levels can reduce (HDL raising) in time.Group (d) can show the reduction of minimum triglyceride, LDL and cholesterol levels.

For atheromatous plaque, group (b), (c) and (e) can show significantly reduced atherosclerotic lesion size in aortal all regions.In group (a) and (d), expection atherosclerotic lesion size does not significantly reduce.

Also expect and only have the mice accepting independent 10g nicotinic acid/kg diet to show cutaneous vasodilation significantly higher compared with every other group that comprises matched group.Compared with group (e), can find that in group (a) to (d) cutaneous vasodilation is lower.

In a word, the dosage used together with 24g leucine/kg diet be the nicotinic acid of 250mg/kg diet to the triglyceride reduced in mice, LDL and cholesterol levels and improve HDL level and can show effect similar compared with independent 10g nicotinic acid/kg diet, and significantly can not increase cutaneous vasodilation.The nicotinic acid of the low dosage used together with resveratrol with leucine is expected and is shown similar effect.

embodiment 5: nicotinic acid/leucine and nicotinic acid/leucine/resveratrol in human body to triglyceride, the impact of LDL, HDL and cholesterol levels

In order to assess effect of title compound, use the title compound comprising (a) nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite and (b) leucine and/or leucine metabolite as herein described to people, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.Said composition can be used to the triglyceride, LDL and the cholesterol levels that reduce after using said composition to people in this people.Be investigated the compositions comprising resveratrol further.

The patient being diagnosed as hyperlipemia is in advance received to carry out randomized double-blind study.The 50mg nicotinic acid that every patient oral (a) is independent, (b) 50mg nicotinic acid and 1135mg leucine, (c) 50mg nicotinic acid, 1135mg leucine and 50mg resveratrol, (d) 1135mg leucine and 50mg resveratrol, e 3000mg nicotinic acid that () is independent, and (f) placebo.Therapeutant every day Orally administered twice, continuous 60 days.

In all groups, the blood sample of patient obtains after application on the the 0th, 30 and 60 day.Measure the serum/plasma level of triglyceride, cholesterol, LDL and HDL.With the uncomfortable horizontal survey cutaneous vasodilation that Laser-Doppler effusion meter and patient describe.

Result

For treatment group, group (a) can demonstrate similar triglyceride, LDL, cholesterol and HDL level with (f) compared with the value of the 0th day, does not have statistically evident difference.Compared with the value of respective 0th day, group (b), (c) and (e) can show remarkable lower triglyceride, LDL and cholesterol levels and significantly higher HDL level in blood flow.Group (d) can show the reduction of minimum triglyceride, LDL and cholesterol levels.

Also expection only has the patient accepting independent 3000mg nicotinic acid just to show cutaneous vasodilation significantly higher compared with every other group that comprises placebo group and the more main suit from patient.Compared with group (e), in group (a) to (d), cutaneous vasodilation can be lower.

In a word, the nicotinic acid of the 50mg dosage used together with 1135mg leucine is to the triglyceride reduced in patient, LDL and cholesterol levels and improve HDL level and can show effect similar compared with independent 3g nicotinic acid, and does not significantly increase cutaneous vasodilation.50mg nicotinic acid+1135mg the leucine used together with 50mg resveratrol can show similar effect.

embodiment 6: nicotinic acid/leucine and nicotinic acid/leucine/resveratrol in human body to Atherosclerosis the impact of dissipating rashes block size

In order to assess effect of title compound, use title compound as herein described to people, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.Said composition can be used to the atheromatous plaque size reduced after using said composition to people in this people.Be investigated the compositions comprising resveratrol further.

The patient experiencing acute chest pain and be diagnosed as hyperlipemia is in advance received to carry out randomized double-blind study.The 50mg nicotinic acid that every patient oral (a) is independent, (b) 50mg nicotinic acid and 1135mg leucine, (c) 50mg nicotinic acid, 1135mg leucine and 50mg resveratrol, (d) 1135mg leucine and 50mg resveratrol, and (e) placebo.Therapeutant every day Orally administered twice, continuous 3 years.At the 0th day, the 6th, 12,18,24,30 and measure the size of atheromatous plaque by Quantitative Coronary Angiography art in 36 months.

Result

For treatment group, group (a) can demonstrate similar atherosclerotic lesion size with (e) compared with the value of the 0th day, and does not have statistically evident difference.Compared with the value of respective 0th day, group (b) and (c) can show significantly reduced atheromatous plaque size.Group (d) can show the reduction of minimum atheromatous plaque size.

embodiment 7: nicotinic acid/leucine is in vivo to triglyceride, LDL, HDL and cholesterol levels impact

In order to assess effect of title compound, use (a) nicotinic acid and (b) leucic title compound of comprising as herein described to mice, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.Said composition reduces triglyceride, LDL and cholesterol levels in mice after using said composition to mice.

Ldl receptor knocks out (LDLRKO) mice is available from JacksonLaboratories (BarHarbor, ME), and grouping is raised under the environment and regular light dark period of room temperature, humid control.Mice is freely obtained before treatment containing 0.21% (weight) cholesterol and the atherogenicity western diet (WD) 4 weeks from 40% calorie of fat and water.For process, give the WD diet that mice (a) is independent, (b) WD and 24g leucine/kg diet, (c) WD and 24g leucine/kg diet and 50mg nicotinic acid/kg diet, (d) WD and 24g leucine/kg diet and 250mg nicotinic acid/kg diet, or (e) WD and 1000mg nicotinic acid/kg diet; This equals the nicotinic acid (about 1,500mg/ day) of the low therapeutic dose in HypercholesterolemiaPatients Patients approx.Process continuous administration 28 days.

The blood sample of all groups of small mouses obtains from the tail of mice all after application on the 28th day.Measure the serum/plasma level of triglyceride, T-CHOL and cholesteryl ester.

Result

Atherogenic western diet causes the remarkable rising of plasma cholesterol (Fig. 5), cholesteryl ester (Fig. 6) and triglyceride (Fig. 7).The interpolation of the nicotinic acid (1,000mg/kg diet) of therapeutic dose causes T-CHOL to reduce by 20% (p<0.01, Fig. 5).Although leucine does not independently affect T-CHOL, but the nicotinic acid (50 or 250mg/kg diet) to sub-therapeutic dose adds leucine to be caused reducing (p<0.01, Fig. 5) with the suitable T-CHOL adopting therapeutic dose to find.Similarly, the nicotinic acid (1 of therapeutic dose, 000mg/kg diet) interpolation cause cholesteryl ester reduce by 28% (p<0.002, Fig. 6), and when adding leucine to the nicotinic acid (50 or 250mg/kg diet) of sub-therapeutic dose, find reduction (p<0.002, Fig. 6) suitable statistically.Leucine does not independently affect cholesteryl ester.Plasma triglyceride is subject to similar impact.The nicotinic acid (1 of therapeutic dose, 000mg/kg diet) interpolation cause plasma triglyceride reduce by 32% (p<0.01, Fig. 7), and the nicotinic acid (50 or 250mg/kg diet) to sub-therapeutic dose has found when adding leucine that triglyceride suitable statistically reduces (p<0.01, Fig. 7).

embodiment 8: leucine-nicotinic acid is on the impact in the life-span of Caenorhabditis elegans

Have studied the purposes comprising (a) nicotinic acid and (b) leucic compositions as described herein, wherein said composition comprises the nicotinic acid of free leucine and sub-therapeutic dose.After using said composition to experimenter, said composition extends the life-span of experimenter synergistically.

Anthelmintic (N2Bristol wild type) available from the hidden rhabditida hereditism center (CaenorhabditisGeneticsCenter) (CGC) of University of Minnesota (UniversityofMinnesota), and using escherichia coli (OP50) on the standard NGM plate of food source in 20 DEG C of growths.Make ovum incubated overnight on the flat board of hunger.Then synchronized L1 larva is transferred to containing shown handled thing, with escherichia coli raise NGM plate on about 35 hours, with reach L4/ children the manhood.In order to study the life-span, 50 young adult worms being placed in inoculation has (the 1st day of=research) on the NGM agar plate of coli strain OP-50.All handled things are all added into agar plate with shown concentration.Handled thing comprises 10nM nicotinic acid and 0.5mM leucine.

Anthelmintic maintains 21 DEG C within the whole persistent period of this research.Every day anthelmintic is transferred to new flat board to eliminate filial generation.If anthelmintic pair does not react with repeatedly contacting of platinum spoon (platinumpick), then marked as dead.Continue this research animal dead to the last.Use Prism6 (GraphPadSoftware) through Kaplan-Meier survival curve analytical data, and determine statistical significance by logarithm order (Log-rank) (Mantel-Cox) inspection.

Find that leucine (0.5mM) and nicotinic acid (10nM) independently do not affect the life-span separately, but upon combination, under primary condition, extend maximum life, and make median life span extend 28% (Fig. 8) under using oxidative stress condition that N,N'-dimethyl-.gamma..gamma.'-dipyridylium (paraquat) (0.2mM) induce.

Although shown at this and described the preferred embodiments of the invention, it will be apparent for a person skilled in the art that this kind of embodiment only provides by way of example.In the case of without departing from the present invention, those skilled in the art will expect many changes, change and replacement now.Should be appreciated that the various replacement schemes of embodiment of the present invention described herein may be used for implementing the present invention.Intention is, limits scope of the present invention with following claim, and the method and structure contained thus in these claim and equivalent thereof.

Claims

1. a compositions, it comprises:

A. at least about 250mg leucine and/or at least about one or more leucine metabolite of 25mg; With

B. at least about 1mg nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite.

2. compositions as claimed in claim 1, wherein said composition is substantially free of nicotiamide.

3. compositions as claimed in claim 1, wherein said composition is substantially free of nicotinic acid metabolite.

4. compositions as claimed in claim 1, wherein said composition is substantially free of each in nicotinoyl coenzyme A, nitocinoylglycine, NAMN, nicotinate adenine dinucleotide and nicotinamide adenine dinucleotide.

5. compositions as claimed in claim 1, the component (b) wherein in said composition is nicotinic acid.

6. compositions as claimed in claim 1, the component (a) wherein in said composition is leucine.

7. compositions as claimed in claim 1, wherein the amount of leucine and/or one or more leucine metabolite is less than about 1g.

8. compositions as claimed in claim 1, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is less than about 1g.

9. compositions as claimed in claim 1, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is less than about 250mg.

10. compositions as claimed in claim 1, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is about 1-100mg.

11. compositionss as claimed in claim 1, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize being less than the nicotinic acid of about 100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

12. compositionss as claimed in claim 1, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize the nicotinic acid of about 10nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

13. compositionss as claimed in claim 1, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize the nicotinic acid of about 1-100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

14. compositionss as claimed in claim 1, wherein said composition makes the triglyceride levels of experimenter, T-CHOL or LDL level reduce at least about 5% effectively.

15. compositionss as claimed in claim 1, wherein the amount of component (a) and (b) reduces the lipid content of described experimenter synergistically when being applied to experimenter.

16. compositionss as claimed in claim 1, the increase that the reduction that the body weight that wherein component (a) and (b) strengthen experimenter synergistically increases, the increase of fat oxidation of experimenter or the Sirt1 of experimenter activate.

17. compositionss as claimed in claim 1, wherein the described amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is not enough to when there is not described leucine and/or one or more leucine metabolite reduce lipid content.

18. compositionss as claimed in claim 1, wherein said composition is included in food.

19. compositionss as claimed in claim 1, a part for wherein said leucine and/or one or more leucine metabolite is free form.

20. compositionss as claimed in claim 1, a part for wherein said leucine and/or one or more leucine metabolite is salt form.

21. compositionss as claimed in claim 1, wherein said composition comprises resveratrol further.

22. compositionss as claimed in claim 1, wherein said composition is formulated for oral administration.

23. compositionss as claimed in claim 1, wherein said composition be tablet, capsule, pill, granule, Emulsion, gel, multiple beadlet, powder, suspension, liquid, semiliquid, semisolid, syrup, serosity or the chewable form be encapsulated in capsule.

24. compositionss as claimed in claim 1, wherein component (a) and component (b) separately packaging.

25. compositionss as claimed in claim 1, wherein component (a) and component (b) are mixing.

26. compositionss as claimed in claim 1, wherein said composition is substantially free of the every seed amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine.

27. compositionss as claimed in claim 1, wherein said composition is substantially free of the often kind of free amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine.

28. compositionss as claimed in claim 1, wherein said composition contains the often kind of free amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine being less than about 0.1%.

29. compositionss as claimed in claim 1, wherein said composition contains the non-leucine aminoacid being less than about 10%.

30. compositionss as claimed in claim 1, one or more leucine metabolite wherein said are selected from ketone-isocaproic acid (KIC), alpha-hydroxy-isocaproic acid and HMB.

31. compositions as claimed in claim 1, wherein said composition not niacinamide-containings.

32. compositionss as claimed in claim 1, it comprises the therapeutic agent that one or more can reduce lipid accumulation further.

33. compositionss as claimed in claim 32, one or more therapeutic agents wherein said are selected from HMG-CoA inhibitor, fibrate, bile acid chelating agent, ezetimibe, his group of Lome, plant sterol, CETP antagonist, orlistat and combination in any thereof.

34. compositionss as claimed in claim 1, the component (a) in wherein said compositions is greater than about 20 with the mol ratio of component (b).

35. compositionss as claimed in claim 1, wherein said composition is configured to unit dosage forms.

36. 1 kinds of test kits, it comprises supplies in many days of the unit dose of the compositions according to any one of claim 0-0 and instructs the description used within a period of time of many days and supply for described many days.

The method of the total cholesterol level in 37. 1 kinds of reduction experimenters in need, it comprises uses compositions as claimed in claim 1 to realize the total cholesterol level in this experimenter to described experimenter.

The method of the total lipid content in 38. 1 kinds of reduction experimenters in need, it comprises uses compositions as claimed in claim 1 to realize the total lipid content in this experimenter to described experimenter.

39. 1 kinds of compositionss, it comprises:

B. a certain amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite,

Wherein said composition is substantially free of each in alanine, glycine, glutamic acid and proline.

40. 1 kinds of compositionss, it comprises:

B. a certain amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite,

Wherein when there is not component (a), the described quantity not sufficient of nicotinic acid and/or nicotinic acid metabolite is to reduce lipid content.

41. 1 kinds of compositionss, it comprises:

A. a certain amount of leucine and/or one or more leucine metabolite; With

Wherein use said composition to experimenter in need, further wherein compared to the dosage of only nicotinic acid with said composition in the lipid content reducing experimenter with identical effectiveness, the cutaneous vasodilation that said composition causes degree to reduce in used experimenter.

42. 1 kinds of compositionss, it comprises:

A. leucine and/or one or more leucine metabolite; With

B. at least about 1mg nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite,

Component (a) in wherein said compositions is greater than about 20 with the mol ratio of component (b).

43. compositionss as described in claim 0-0, wherein said composition comprises at least about 500mg leucine and/or at least about one or more leucine metabolite described in 200mg.

44. compositionss as described in claim 0-0, wherein said composition comprises at least about 250mg leucine and/or one or more leucine metabolite of about 25mg.

45. compositionss as described in claim 39-42, wherein said composition is substantially free of nicotiamide.

46. compositionss as described in claim 39-42, wherein said composition is substantially free of nicotinic acid metabolite.

47. compositionss as described in claim 39-42, wherein said composition is substantially free of each in nicotinoyl coenzyme A, nitocinoylglycine, NAMN, nicotinate adenine dinucleotide and nicotinamide adenine dinucleotide.

48. compositionss as described in claim 39-42, the component (b) wherein in said composition is nicotinic acid.

49. compositionss as described in claim 39-42, the component (a) wherein in said composition is leucine.

50. compositionss as described in claim 39-42, wherein the amount of leucine and/or one or more leucine metabolite is less than about 1g.

51. compositionss as described in claim 39-42, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is less than about 1g.

52. compositionss as described in claim 39-42, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is less than about 250mg.

53. compositionss as described in claim 39-42, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is about 1-100mg.

54. compositionss as described in claim 39-41, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is at least about 1mg.

Compositions as described in claim 39-42, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize being less than the nicotinic acid of about 100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

55. compositionss as described in claim 39-42, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize the nicotinic acid of about 10nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

56. compositionss as described in claim 39-42, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize the nicotinic acid of about 1-100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

57. compositionss as described in claim 39-42, wherein said composition makes the triglyceride levels of experimenter, T-CHOL or LDL level reduce at least about 5% effectively.

58. compositionss as described in claim 39-42, wherein the amount of component (a) and (b) reduces the lipid content of described experimenter synergistically when being applied to experimenter.

59. compositionss as described in claim 39-42, the increase that the reduction that the body weight that wherein component (a) and component (b) strengthen experimenter synergistically increases, the increase of fat oxidation of experimenter or the Sirt1 of experimenter activate.

60. compositionss as described in claim 39-42, wherein when there is not described leucine and/or one or more leucine metabolite, the quantity not sufficient of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is to reduce lipid content.

61. compositionss as described in claim 39-42, wherein said composition is included in food.

62. compositionss as described in claim 39-42, a part for wherein said leucine and/or one or more leucine metabolite is free form.

63. compositionss as described in claim 39-42, a part for wherein said leucine and/or one or more leucine metabolite is salt form.

64. compositionss as described in claim 39-42, wherein said composition comprises resveratrol further.

65. compositionss as described in claim 39-42, wherein said composition is formulated for oral administration.

66. compositionss as described in claim 39-42, wherein said composition be tablet, capsule, pill, granule, Emulsion, gel, multiple beadlet, powder, suspension, liquid, semiliquid, semisolid, syrup, serosity or the chewable form be encapsulated in capsule.

67. compositionss as described in claim 39-42, wherein component (a) and component (b) separately packaging.

68. compositionss as described in claim 39-42, wherein component (a) and component (b) are mixing.

69. compositionss as described in claim 39-42, wherein said composition is substantially free of the every seed amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine.

70. compositionss as described in claim 39-42, wherein said composition is substantially free of the often kind of free amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine.

71. compositionss as described in claim 39-42, wherein said composition contains the often kind of free amino acid being selected from alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine, isoleucine and tyrosine being less than about 0.1%.

72. compositionss as described in claim 39-42, wherein said composition contains the non-leucine aminoacid being less than about 10%.

73. compositionss as described in claim 39-42, one or more leucine metabolite wherein said are selected from ketone-isocaproic acid (KIC), alpha-hydroxy-isocaproic acid and HMB.

74. compositionss as described in claim 39-42, wherein said composition not niacinamide-containing.

75. compositionss as described in claim 39-42, it comprises the therapeutic agent that one or more can reduce lipid accumulation further.

76. compositionss as described in claim 75, one or more therapeutic agents wherein said are selected from HMG-CoA inhibitor, fibrate, bile acid chelating agent, ezetimibe, his group of Lome, plant sterol, CETP antagonist, orlistat and combination in any thereof.

77. compositionss as described in claim 39-42, the component (a) in wherein said compositions is greater than about 20 with the mol ratio of (b).

78. compositionss as described in claim 39-42, wherein said composition is configured to unit dosage forms.

79. 1 kinds of test kits, it comprises supplies in many days of the unit dose of the compositions according to any one of claim 39-78 and instructs the description used within a period of time of many days and supply for described many days.

80. 1 kinds of methods reducing the total cholesterol level in experimenters in need, it comprises uses compositions as described in claim 39-42 to realize the total cholesterol level in this experimenter to described experimenter.

81. 1 kinds of methods reducing the total lipid content in experimenters in need, it comprises uses compositions as described in claim 39-42 to realize the total lipid content in this experimenter to described experimenter.

The method of the total cholesterol level in 82. 1 kinds of reduction experimenters in need, it comprises the compositions comprising leucine and/or one or more leucine metabolite and a certain amount of nicotinic acid and/or nicotinic acid metabolite using doses to described experimenter, to realize the total cholesterol level in this experimenter.

83. methods as described in claim 82, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is less than about 250mg.

84. methods as described in claim 82, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is less than about 100mg.

85. methods as described in claim 82, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is less than about 25mg.

86. methods as described in claim 82, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is less than about 10mg.

87. methods as described in claim 82, wherein said dosage is unit dose.

88. 1 kinds of methods reducing the side effect of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite, wherein the feature of this side effect is the increase of the cutaneous vasodilation of the experimenter using nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite described, and the method comprises the compositions using leucine and/or one or more leucine metabolite including effective amount to the described experimenter using nicotinic acid and/or one or more nicotinic acid metabolite.

89. methods as described in claim 88, wherein said compositions oral administration.

90. methods as described in claim 88, wherein said effective dose comprises at least about 500mg leucine and/or at least about one or more leucine metabolite described in 200mg.

91. methods as described in claim 88, wherein said effective dose comprises at least about 250mg leucine and/or at least about one or more leucine metabolite described in 25mg.

92. methods as described in claim 88, if be sub-therapeutic dose when wherein nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite are used separately.

93. methods as described in claim 88, wherein nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite are the amount being less than about 1g.

94. methods as described in claim 88, wherein nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite are the amount being less than about 250mg.

95. methods as described in claim 88, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite is about 1-100mg.

96. compositionss as described in claim 88, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or one or more nicotinic acid metabolite can realize the nicotinic acid of about 1-100nM and/or the serum levels of nicotinamide riboside and/or one or more nicotinic acid metabolite.

The method of the atheromatous plaque size in 97. 1 kinds of reduction experimenters in need, it comprises the compositions comprising leucine and/or one or more leucine metabolite and a certain amount of nicotinic acid and/or nicotinic acid metabolite using doses to described experimenter, to realize the total atheromatous plaque size in experimenter.

98. methods as described in claim 97, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is less than about 250mg.

99. methods as described in claim 970, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is about 1-100mg.

100. methods as described in claim 97, wherein the amount of nicotinic acid and/or nicotinamide riboside and/or nicotinic acid metabolite is less than about 25mg.

101. methods as described in claim 97, wherein to described experimenter's applying said compositions at least about 1 year.

102. methods as described in claim 97, wherein said dosage is unit dose.